Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Takeshi Uchiumi Last modified date:2019.05.31

Professor / Department of Health Sciences, / Department of Health Sciences / Faculty of Medical Sciences


Papers
1. Masako Ichiyama, Hirosuke Inoue, Masayuki Ochiai, masataka ishimura, Akira Shiraishi, Junko Fujiyoshi, Hironori Yamashita, Kazuo Sato, Shinya Matsumoto, Taeko Hotta, Takeshi Uchiumi, Dongchon Kang, Shoichi Ohga, Diagnostic challenge of the newborn patients with heritable protein C deficiency, Journal of Perinatology, 10.1038/s41372-018-0262-0, 39, 2, 212-219, 2019.02, Objective: The diagnosis of neonatal-onset protein C (PC) deficiency is challenging. This study aimed to establish the neonatal screening of heritable PC deficiency in Japan. Study design: We determined the changes in plasma activity levels of PC and protein S (PS) in healthy neonates, and studied newborn patients with PROC mutation in the Japanese registry. Result: Physiological PC and PS levels increased with wide range. The PC/PS-activity ratios converged after birth. The PC/PS-activity ratios of 19 patients with biallelic mutations, but not, 9 with monoallelic mutation, were lower than those of 13 without mutation. The logistic regression analyses established a formula including two significant variables of PC activity (cut-off < 10%, odds ratio = 30.0) and PC/PS-activity ratio (cut-off < 0.35, odds ratio = 22.7), with 93% sensitivity and 44% specificity for determining patients with mutation(s). Conclusion: The PC/PS-activity ratio is an effective parameter for the genetic screening of neonatal-onset PC-deficiency in Japanese population..
2. masaki shiota, Naohiro Fujimoto, Shigehiro Tsukahara, Miho Ushijima, ario takeuchi, Eiji Kashiwagi, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Masatoshi Eto, Genetic Polymorphism in Sex Hormone-binding Globulin With a Prognosis of Androgen Deprivation Therapy in Metastatic Prostate Cancer Among Japanese Men, Clinical Genitourinary Cancer, 10.1016/j.clgc.2019.03.021, 2019.01, This study investigated the impact of a missense polymorphism (rs6259, D356N)in the SHBG gene among 104 Japanese treated with primary androgen deprivation therapy (ADT)for metastatic prostate cancer. Genotype in SHBG was associated with progression and any-cause death, but not with serum testosterone levels during ADT. This study suggested SHBG variation might be an independent prognostic biomarker in ADT..
3. masaki shiota, Naohiro Fujimoto, Shigehiro Tsukahara, Miho Ushijima, ario takeuchi, Eiji Kashiwagi, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Masatoshi Eto, The impact of genetic polymorphism on CYP19A1 in androgen-deprivation therapy among Japanese men, Cancer chemotherapy and pharmacology, 10.1007/s00280-019-03811-8, 2019.01, Purpose: Inadequate suppression of testosterone during androgen-deprivation therapy impairs its efficacy. This study investigated the significance of genetic polymorphism in CYP19A1, which encodes aromatase that catalyzes androgens into estrogens, among men treated with primary ADT for metastatic prostate cancer. Methods: This study included 80 Japanese patients with metastatic prostate cancer whose serum testosterone levels during ADT were available. The association of CYP19A1 gene polymorphism (rs1870050) with clinicopathological parameters including serum testosterone levels during ADT as well as progression-free survival and overall survival was examined. Results: Serum testosterone levels during ADT of men carrying homozygous wild-type (AA) in the CYP19A1 gene [median (interquartile range); 11.6 (8.3–20.3) ng/dl] were higher than those in men carrying the heterozygous/homozygous variant (AC/CC) [median (interquartile range); 10.0 (6.4–12.8) ng/dl]. When adjusted by Gleason score, initial PSA, M-stage and serum testosterone level during ADT, heterozygous/homozygous variant (AC/CC) in the CYP19A1 gene was associated with a lower risk of progression to castration resistance [hazard ratio (95% confidence interval), 0.53 [0.29–0.92], p = 0.025], but not to any-cause death [hazard ratio (95% confidence interval), 0.74 [0.36–1.49], p = 0.40]. Conclusions: These findings suggest that genetic variation in CYP19A1 (rs1870050) might affect the prognosis of patients with metastatic prostate cancer when treated with ADT by regulating serum testosterone levels..
4. Gotoh K, Morisaki T, Setoyama D, Sasaki K, Yagi M, Igami K, Mizuguchi S, Uchiumi T, Fukui Y and Kang D, Mitochondrial p32/C1qbp Is a Critical Regulator of Dendritic Cell Metabolism and Maturation, CELL REPORTS, 10.1016/j.celrep.2018.10.057, 25, 7, 1800-+, 2018.11, Dendritic cell (DC) maturation induced by Toll-like receptor agonists requires activation of downstream signal transduction and metabolic changes. The endogenous metabolite citrate has recently emerged as a modulator of DC activation. However, the metabolic requirements that support citrate production remain poorly defined. Here, we demonstrate that p32/C1qbp, which functions as a multifunctional chaperone protein in mitochondria, supports mitochondrial metabolism and DC maturation. Metabolic analysis revealed that the citrate increase induced by lipopolysaccharide (LPS) is impaired in p32-deficient DCs. We also found that p32 interacts with dihydrolipoamide S-acetyltransferase (E2 component of pyruvate dehydrogenase [PDH] complex) and positively regulates PDH activity in DCs. Therefore, we suggest that DC maturation is regulated by citrate production via p32-dependent PDH activity. p32-null mice administered a PDH inhibitor show decreased DC maturation and ovalbumin-specific IgG production in vivo, suggesting that p32 may serve as a therapeutic target for DC-related autoimmune diseases..
5. masaki shiota, Takashi Dejima, Yoshiaki Yamamoto, ario takeuchi, Kenjiro Imada, Eiji Kashiwagi, Junichi Inokuchi, Katsunori Tatsugami, Shunichi Kajioka, Takeshi Uchiumi, Masatoshi Eto, Collateral resistance to taxanes in enzalutamide-resistant prostate cancer through aberrant androgen receptor and its variants, Cancer Science, 10.1111/cas.13751, 109, 10, 3224-3234, 2018.10, Currently, the optimal sequential use of androgen receptor (AR) axis-targeted agents and taxane chemotherapies remains undetermined. We aimed to elucidate the resistance status between taxanes and enzalutamide, and the functional role of the AR axis. Enzalutamide-resistant 22Rv1 cells showed collateral resistance to taxanes, including docetaxel and cabazitaxel. However, taxane-resistant cells showed no collateral resistance to enzalutamide; taxane-resistant cells expressed comparable protein levels of full-length AR and AR variants. Knockdown of both full-length AR and AR variants rendered cells sensitive to taxanes, whereas knockdown of AR variants sensitized cells to enzalutamide, but not to taxanes. In contrast, overexpression of full-length AR rendered cells resistant to taxanes. Consistently, the prostate-specific antigen response and progression-free survival in docetaxel chemotherapy were worse in cases with prior use of ARAT agents compared with cases without. Collateral resistance to taxanes was evident after obtaining enzalutamide resistance, and aberrant AR signaling might be involved in taxane resistance..
6. Takuma Hayami, Akihiko Yamaguchi, Takeshi Kato, Toshihiro Tanaka, Yuka Nishizawa, Takahide Yanagi, Takashi Taga, Shinya Matsumoto, Takeshi Uchiumi, Noriki Fujimoto, Purpura fulminans in congenital protein C deficiency successfully treated with fresh frozen plasma and thrombomodulin, Journal of Dermatology, 10.1111/1346-8138.14194, 45, 6, e165-e166, 2018.06.
7. masaki shiota, Naohiro Fujimoto, Katuyoshi Higashijima, Kenjiro Imada, Eiji Kashiwagi, ario takeuchi, Junichi Inokuchi, Katsunori Tatsugami, Shunichi Kajioka, Takeshi Uchiumi, Masatoshi Eto, Mineralocorticoid receptor signaling affects therapeutic effect of enzalutamide, Prostate, 10.1002/pros.23661, 2018.01, Background: Corticosteroids play important roles in prostate cancer therapeutics. However, their role when combined with enzalutamide remains obscure. Then, we aimed to elucidate the functional and clinical impact of corticosteroids on steroid receptors in androgen receptor (AR)-targeting therapy utilizing enzalutamide. Methods: The therapeutic effect was studied according to concomitant use of corticosteroids in 86 men treated with enzalutamide. The sensitivity to various agents was evaluated using cytotoxicity assays in prostate cancer cells. Gene expression levels were evaluated by quantitative real-time polymerase chain reaction in prostate cancer cells and tissues. Results: The therapeutic effect of enzalutamide was particularly lessened with concomitant treatment with dexamethasone. Consistently, dexamethasone increased cellular resistance to enzalutamide while prednisolone and aldosterone decreased cellular resistance to enzalutamide in prostate cancer cells. Inversely, mineralocorticoid receptor (MR) knockdown augmented the activity of AR signaling and the cellular resistance to enzalutamide. Conclusions: MR plays a critical role in resistance to AR-targeting therapies, which may be overcome by activation of MR signaling..
8. masaki shiota, Naohiro Fujimoto, ario takeuchi, Eiji Kashiwagi, Takashi Dejima, Junichi Inokuchi, Katsunori Tatsugami, Akira Yokomizo, Shunichi Kajioka, Takeshi Uchiumi, Masatoshi Eto, The Association of Polymorphisms in the Gene Encoding Gonadotropin-Releasing Hormone with Serum Testosterone Level during Androgen Deprivation Therapy and Prognosis of Metastatic Prostate Cancer, Journal of Urology, 10.1016/j.juro.2017.09.076, 2018.01, Purpose: Serum testosterone suppression during androgen deprivation therapy has been reported to affect the efficacy of androgen deprivation therapy. However, the factors impacting hormonal variations during androgen deprivation therapy remain unclear. Therefore, in this study we investigated the significance of missense polymorphisms in the gene encoding GNRH in men treated with primary androgen deprivation therapy for metastatic prostate cancer. Materials and Methods: This study included 80 Japanese patients with metastatic prostate cancer with available serum testosterone levels during androgen deprivation therapy. We examined the association of GNRH1 (rs6185, S20W) and GNRH2 (rs6051545, A16V) gene polymorphisms with clinicopathological parameters, including serum testosterone levels during androgen deprivation therapy, as well as prognosis, including progression-free and overall survival. Results: The CT and CT/TT alleles in the GNRH2 gene (rs6051545) were associated with higher serum testosterone during androgen deprivation therapy compared with those of the CC allele. Consequently the CT alleles were associated with a higher risk of progression after adjustment for age and serum testosterone during androgen deprivation therapy (HR 1.73, 95% CI 1.00-3.00, p = 0.049). Conclusions: Taken together these findings suggest that rs6051545 (GNRH2) genetic variation may result in inadequate suppression of serum testosterone during androgen deprivation therapy. This may lead to detrimental effects of androgen deprivation therapy on prognosis in men with metastatic prostate cancer..
9. masaki shiota, Naohiro Fujimoto, Kenjiro Imada, Eiji Kashiwagi, ario takeuchi, Junichi Inokuchi, Katsunori Tatsugami, Shunichi Kajioka, Takeshi Uchiumi, Masatoshi Eto, Prognostic impact of genetic polymorphism in mineralocorticoid receptor and comorbidity with hypertension in androgen-deprivation therapy, Frontiers in Oncology, 10.3389/fonc.2018.00635, 8, DEC, 2018.01, Mineralocorticoid receptor (MR) signaling which is closely associated with hypertension plays important roles in resistance to antiandrogen therapy in prostate cancer. However, its impact on the prognosis in androgen-deprivation therapy (ADT) has not been elucidated. Then, we investigated the impact of genetic variation in MR and comorbidity with hypertension on the prognosis in ADT. This study included 182 Japanese patients with prostate cancer treated with ADT, whose comorbidity status with hypertension were available. The associations of MR polymorphism (rs5522) and comorbidity with hypertension with clinicopathological parameters as well as progression-free survival and overall survival were examined. Clinicopathological characteristics were comparable between genetic variation in MR. However, homozygous variant in MR was associated with shorter time to castration resistance (P = 0.014) and any-cause death (P = 0.024). In patients' background, presence of comorbidity with hypertension showed the trend with lower PSA level at diagnosis and lower biopsy Gleason score, as well as significant association with less incidence of N1. Comorbidity with hypertension was associated with longer time to castration resistance (P = 0.043) and any-cause death (P = 0.046), which was diminished on multivariate analysis including age, PSA level at diagnosis, biopsy Gleason score, clinical stage, and the modality of hormonal therapy. Genetic variation in MR (rs5522) and comorbidity with hypertension were significantly and potentially associated with prognosis when treated with ADT, respectively. This suggests that the individual intensity of MR signaling may be associated with resistance to ADT and a promising biomarker in ADT..
10. Yuichi Matsushima, Yuta Hirofuji, Masamune Aihara, Song Yue, Takeshi Uchiumi, Laurie S. Kaguni, Dongchon Kang, Drosophila protease ClpXP specifically degrades DmLRPPRC1 controlling mitochondrial mRNA and translation, Scientific Reports, 10.1038/s41598-017-08088-6, 7, 1, 2017.12, ClpXP is the major protease in the mitochondrial matrix in eukaryotes, and is well conserved among species. ClpXP is composed of a proteolytic subunit, ClpP, and a chaperone-like subunit, ClpX. Although it has been proposed that ClpXP is required for the mitochondrial unfolded protein response, additional roles for ClpXP in mitochondrial biogenesis are unclear. Here, we found that Drosophila leucine-rich pentatricopeptide repeat domain-containing protein 1 (DmLRPPRC1) is a specific substrate of ClpXP. Depletion or introduction of catalytically inactive mutation of ClpP increases DmLRPPRC1 and causes non-uniform increases of mitochondrial mRNAs, accumulation of some unprocessed mitochondrial transcripts, and modest repression of mitochondrial translation in Drosophila Schneider S2 cells. Moreover, DmLRPPRC1 over-expression induces the phenotypes similar to those observed when ClpP is depleted. Taken together, ClpXP regulates mitochondrial gene expression by changing the protein level of DmLRPPRC1 in Drosophila Schneider S2 cells..
11. Satomi Mezuki, Kenji Fukuda, Tomonaga Matsushita, Yoshihisa Fukushima, ryu matsuo, Yu ichi Goto, Takehiro Yasukawa, Takeshi Uchiumi, Dongchon Kang, Takanari Kitazono, Tetsuro Ago, Isolated and repeated stroke-like episodes in a middle-aged man with a mitochondrial ND3 T10158C mutation
A case report, BMC Neurology, 10.1186/s12883-017-1001-4, 17, 1, 2017.12, Background: Mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) syndrome, is the most common phenotype of mitochondrial disease. It often develops in childhood or adolescence, usually before the age of 40, in a maternally-inherited manner. Mutations in mitochondrial DNA (mtDNA) are frequently responsible for MELAS. Case presentation: A 55-year-old man, who had no family or past history of mitochondrial disorders, suddenly developed bilateral visual field constriction and repeated stroke-like episodes. He ultimately presented with cortical blindness, recurrent epilepsy and severe cognitive impairment approximately 6 months after the first episode. Genetic analysis of biopsied biceps brachii muscle, but not of peripheral white blood cells, revealed a T10158C mutation in the mtDNA-encoded gene of NADH dehydrogenase subunit 3 (ND3), which has previously been thought to be associated with severe or fatal mitochondrial disorders that develop during the neonatal period or in infancy. Conclusion: A T10158C mutation in the ND3 gene can cause atypical adult-onset stroke-like episodes in a sporadic manner..
12. Mikako Yagi, Takeshi Uchiumi, Noriaki Sagata, Daiki Setoyama, Rie Amamoto, Yuichi Matsushima, Dongchon Kang, Neural-specific deletion of mitochondrial p32/C1qbp leads to leukoencephalopathy due to undifferentiated oligodendrocyte and axon degeneration, Scientific Reports, 10.1038/s41598-017-15414-5, 7, 1, 2017.12, Mitochondrial dysfunction is a critical step in the pathogenesis of many neurodegenerative diseases. The p32/ C1qbp gene functions as an essential RNA and protein chaperone in mitochondrial translation, and is indispensable for embryonic development. However, little is known about the consequences of mitochondrial dysfunction of p32 deletion in the brain development. Here, we found that mice lacking p32 in the central nervous system (p32cKO mice) showed white matter degeneration accompanied by progressive oligodendrocyte loss, axon degeneration and vacuolation in the mid brain and brain stem regions. Furthermore, p32cKO mice died within 8 weeks of birth. We also found that p32-deficient oligodendrocytes and neurons showed reduced oligodendrocyte differentiation and axon degeneration in primary culture. We show that mitochondrial disruption activates an adaptive program known as the integrated stress response (ISR). Mitochondrial respiratory chain function in oligodendrocytes and neurons is, therefore, essential for myelination and axon maintenance, respectively, suggesting that mitochondrial respiratory chain dysfunction in the central nervous system contributes to leukoencephalopathy..
13. Kensuke Yamamichi, Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Yuki Nakao, Karen Yotsumoto, Hiroaki Yamato, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Amelogenin induces M2 macrophage polarisation via PGE2/cAMP signalling pathway, Archives of Oral Biology, 10.1016/j.archoralbio.2017.08.005, 83, 241-251, 2017.11, Objectives Amelogenin, the major component of the enamel matrix derivative (EMD), has been suggested as a bioactive candidate for periodontal regeneration. Apart from producing a regenerative effect on periodontal tissues, amelogenin has also been reported to have an anti-inflammatory effect. However, the precise molecular mechanisms underlying these effects remain unclear. In the present study, we examined the immunomodulatory effects of amelogenin on macrophages. Design Human phorbol 12-myristate 13-acetate (PMA)-differentiated U937 macrophages and CD14+ peripheral blood-derived monocytes (PBMC)-derived macrophages were stimulated with recombinant amelogenin (rM180). After performing a detailed microarray analysis, the effects of rM180 on macrophage phenotype and signal transduction pathways were evaluated by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, confocal microscopy and flow cytometry. Results The microarray analysis demonstrated that rM180 increased the expression of anti-inflammatory genes in lipopolysaccharide (LPS)-challenged macrophages after 24 h, while it temporarily up-regulated inflammatory responses at 4 h. rM180 significantly enhanced the expression of M2 macrophage markers (CD163 and CD206). rM180-induced M2 macrophage polarisation was associated with morphological changes as well as vascular endothelial growth factor (VEGF) production. rM180 enhanced prostaglandin E2 (PGE2) expression, and the activation of the cAMP/cAMP-responsive element binding (CREB) signaling pathway was involved in amelogenin-induced M2 macrophage polarisation. Blocking of PGE2 signaling by indomethacin specifically abrogated rM180 with or without LPS-induced M2 shift in PBMC-derived macrophages. Conclusion Amelogenin could reprogram macrophages into the anti-inflammatory M2 phenotype. It could therefore contribute to the early resolution of inflammation in periodontal lesions and provide a suitable environment for remodeling-periodontal tissues..
14. Takashi Matsumoto, Takeshi Uchiumi, Keisuke Monji, Mikako Yagi, Daiki Setoyama, Rie Amamoto, Yuichi Matsushima, masaki shiota, Masatoshi Eto, Dongchon Kang, Doxycycline induces apoptosis via ER stress selectively to cells with a cancer stem cell-like properties
Importance of stem cell plasticity, Oncogenesis, 10.1038/s41389-017-0009-3, 6, 11, 2017.11, Tumor heterogeneity can be traced back to a small subset of cancer stem cells (CSCs), which can be derived from a single stem cell and show chemoresistance. Recent studies showed that CSCs are sensitive to mitochondrial targeting antibiotics such as doxycycline. However, little is known about how cancer cells undergo sphere formation and how antibiotics inhibit CSC proliferation. Here we show that under sphere-forming assay conditions, prostate cancer cells acquired CSC-like properties: promoted mitochondrial respiratory chain activity, expression of characteristic CSC markers and resistance to anticancer agents. Furthermore, those CSC-like properties could reversibly change depending on the culture conditions, suggesting some kinds of CSCs have plasticity in tumor microenvironments. The sphere-forming cells (i.e. cancer stem-like cells) showed increased contact between mitochondria and mitochondrial associated-endoplasmic reticulum (ER) membranes (MAM). Mitochondrial targeting doxycycline induced activating transcription factor 4 (ATF4) mediated expression of ER stress response and led to p53-upregulated modulator of apoptosis (PUMA)-dependent apoptosis only in the cancer stem-like cells. We also found that doxycycline effectively suppressed the sphere formation in vitro and blocked CD44v9-expressing tumor growth in vivo. In summary, these data provide new molecular findings that monolayer cancer cells acquire CSC-like properties in a reversible manner. These findings provide important insights into CSC biology and a potential new treatment of targeting mitochondria dependency..
15. René G. Feichtinger, Monika Oláhová, Yoshihito Kishita, Caterina Garone, Laura S. Kremer, Mikako Yagi, Takeshi Uchiumi, Alexis A. Jourdain, Kyle Thompson, Aaron R. D'Souza, Robert Kopajtich, Charlotte L. Alston, Johannes Koch, Wolfgang Sperl, Elisa Mastantuono, Tim M. Strom, Saskia B. Wortmann, Thomas Meitinger, Germaine Pierre, Patrick F. Chinnery, Zofia M. Chrzanowska-Lightowlers, Robert N. Lightowlers, Salvatore DiMauro, Sarah E. Calvo, Vamsi K. Mootha, Maurizio Moggio, Monica Sciacco, Giacomo P. Comi, Dario Ronchi, Kei Murayama, Akira Ohtake, Pedro Rebelo-Guiomar, Masakazu Kohda, Dongchon Kang, Johannes A. Mayr, Robert W. Taylor, Yasushi Okazaki, Michal Minczuk, Holger Prokisch, Biallelic C1QBP Mutations Cause Severe Neonatal-, Childhood-, or Later-Onset Cardiomyopathy Associated with Combined Respiratory-Chain Deficiencies, American Journal of Human Genetics, 10.1016/j.ajhg.2017.08.015, 101, 4, 525-538, 2017.10, Complement component 1 Q subcomponent-binding protein (C1QBP; also known as p32) is a multi-compartmental protein whose precise function remains unknown. It is an evolutionary conserved multifunctional protein localized primarily in the mitochondrial matrix and has roles in inflammation and infection processes, mitochondrial ribosome biogenesis, and regulation of apoptosis and nuclear transcription. It has an N-terminal mitochondrial targeting peptide that is proteolytically processed after import into the mitochondrial matrix, where it forms a homotrimeric complex organized in a doughnut-shaped structure. Although C1QBP has been reported to exert pleiotropic effects on many cellular processes, we report here four individuals from unrelated families where biallelic mutations in C1QBP cause a defect in mitochondrial energy metabolism. Infants presented with cardiomyopathy accompanied by multisystemic involvement (liver, kidney, and brain), and children and adults presented with myopathy and progressive external ophthalmoplegia. Multiple mitochondrial respiratory-chain defects, associated with the accumulation of multiple deletions of mitochondrial DNA in the later-onset myopathic cases, were identified in all affected individuals. Steady-state C1QBP levels were decreased in all individuals’ samples, leading to combined respiratory-chain enzyme deficiency of complexes I, III, and IV. C1qbp−/− mouse embryonic fibroblasts (MEFs) resembled the human disease phenotype by showing multiple defects in oxidative phosphorylation (OXPHOS). Complementation with wild-type, but not mutagenized, C1qbp restored OXPHOS protein levels and mitochondrial enzyme activities in C1qbp−/− MEFs. C1QBP deficiency represents an important mitochondrial disorder associated with a clinical spectrum ranging from infantile lactic acidosis to childhood (cardio)myopathy and late-onset progressive external ophthalmoplegia..
16. Toshiro Saito, Takeshi Uchiumi, Mikako Yagi, Rie Amamoto, Daiki Setoyama, Yuichi Matsushima, Dongchon Kang, Cardiomyocyte-specific loss of mitochondrial p32/C1qbp causes cardiomyopathy and activates stress responses, Cardiovascular Research, 10.1093/cvr/cvx095, 113, 10, 1173-1185, 2017.08, Aims Mitochondria are important organelles, dedicated to energy production. Mitochondrial p32/C1qbp, which functions as an RNA and protein chaperone, interacts with mitochondrial mRNA and is indispensable for mitochondrial function through its regulation of mitochondrial translation in cultured cell lines. However, the precise role of p32/C1qbp in vivo is poorly understood because of embryonic lethality in the systemic p32-deficient mouse. The goal of this study was to examine the physiological function of mitochondrial p32/C1qbp in the heart. Methods and results We investigated the role of p32 in regulating cardiac function in mice using a Cre-loxP recombinase technology against p32 with tamoxifen-inducible knockdown or genetic ablation during postnatal periods. Cardiomyocyte-specific deletion of p32 resulted in contractile dysfunction, cardiac dilatation and cardiac fibrosis, compared with hearts of control mice. We also found decreased COX1 expression, decreased rates of oxygen consumption and increased oxidative stress, indicating that these mice had cardiac mitochondrial dysfunction provoked by p32-deficiency at early stage. Next, we investigated lifespan in cardiac-specific p32-deficient mice. The mice died beginning at 12 months and their median lifespan was ∼14 months. Cardiac mitochondria in the p32-deficient mice showed disordered alignment, enlargement and abnormalities in their internal structure by electron microscopy. We observed that, in p32-deficient compared with control myocytes, AMPKI' was constitutively phosphorylated and 4EBP-1 and ribosomal S6K were less phosphorylated, suggesting impairment of mammalian target of rapamycin signalling. Finally, we found that expression levels of mitokines such as FGF21 and of integrated stress response genes were significantly increased. Metabolic analysis demonstrated that the urea cycle was impaired in the p32-deficient hearts. Conclusion These findings support a key role for mitochondrial p32 protein in cardiac myocytes modulating mitochondrial translation and function, and thereby survival..
17. Hirofumi Inoue, Shin Ichi Terachi, Takeshi Uchiumi, Tetsuji Sato, Michiyo Urata, masataka ishimura, Yui Koga, Taeko Hotta, Toshiro Hara, Dongchon Kang, Shoichi Ohga, The clinical presentation and genotype of protein C deficiency with double mutations of the protein C gene, Pediatric Blood and Cancer, 10.1002/pbc.26404, 64, 7, 2017.07, Background: Severe protein C (PC) deficiency is a rare heritable thrombophilia leading to thromboembolic events during the neonatal period. It remains unclear how individuals with complete PC gene (PROC) defects develop or escape neonatal stroke or purpura fulminans (PF). Procedure: We studied the onset of disease and the genotype of 22 PC-deficient patients with double mutations in PROC based on our cohort (n = 12) and the previous reports (n = 10) in Japan. Results: Twenty-two patients in 20 unrelated families had 4 homozygous and 18 compound heterozygous mutations. Sixteen newborns presented with PF (n = 11, 69%), intracranial thromboembolism and hemorrhage (n = 13, 81%), or both (n = 8, 50%), with most showing a plasma PC activity of <10%. Six others first developed overt thromboembolism when they were over 15 years of age, showing a median PC activity of 31% (range: 19–52%). Fifteen of the 22 patients (68%) had the five major mutations (G423VfsX82, V339M, R211W, M406I, and F181V) or two others (E68K and K193del) that have been reported in Japan. Three of the six late-onset cases, but none of the 16 neonatal cases, had the K193del mutation, which has been reported to be the most common variant of Chinese thrombophilia. A novel mutation of A309V was determined in a family of two patients with late onset. Conclusions: The genotype of double-PROC mutants might show less diversity than heterozygous mutants in terms of the timing of the onset of thrombophilia (newborn onset or late onset)..
18. Katsuhiko Sasaki, Kazuhito Gotou, Sho Miake, Daiki Setoyama, Mikako Yagi, Ko Igami, Takeshi Uchiumi, Dongchon Kang, p32 is Required for Appropriate Interleukin-6 Production Upon LPS Stimulation and Protects Mice from Endotoxin Shock, EBioMedicine, 10.1016/j.ebiom.2017.05.018, 20, 161-172, 2017.06, Sepsis is a major cause of morbidity and mortality in seriously ill patients and mitochondrial dysfunction is associated with poor outcomes in septic patients. Although interleukin-6 (IL-6) is a good prognostic marker for sepsis, the relationship between mitochondrial dysfunction and IL-6 remains poorly understood. We identified p32/C1QBP/HABP1 as a regulator of IL-6 production in response to lipopolysaccharide (LPS). LPS induced IL-6 overproduction in p32 deficient mouse embryonic fibroblasts (MEFs) through NF-κB independent but activating transcription factor (ATF) 4 dependent pathways. Short hairpin RNA-based knockdown of ATF4 in p32 deficient MEFs markedly inhibited LPS-induced IL-6 production. Furthermore, MEFs treated with chloramphenicol, an inhibitor of mitochondrial translation, produced excessive IL-6 via ATF4 pathways. Using a LPS-induced endotoxin shock model, mice with p32 ablation in myeloid cells showed increased lethality and overproduction of IL-6. Thus, this study provides a molecular link how mitochondrial dysfunction leads to IL-6 overproduction and poor prognosis of sepsis..
19. masaki shiota, Akira Yokomizo, ario takeuchi, Eiji Kashiwagi, Takashi Dejima, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Masatoshi Eto, Protein kinase C regulates Twist1 expression via NF-κB in prostate cancer, Endocrine-Related Cancer, 10.1530/ERC-16-0384, 24, 4, 171-180, 2017.04, The progression of prostate cancer to metastatic and castration-resistant disease represents a critical step. We previously showed that protein kinase C (PKC) activation followed by Twist1 and androgen receptor (AR) induction played a critical role in castration resistance, but the precise molecular mechanism remains unknown. This study aimed to elucidate the relevant molecular mechanism, focusing on NF-κB transcription factor. We examined the activity of NF-κB after PKC inhibition, and the expression of Twist1 and AR after inhibition of NF-κB in human prostate cancer cells. We also investigated the status of PKC/NF-κB after inhibition of AR signaling in cells resistant to hormonal therapy. As a result, inhibition of PKC signaling using knockdown and smallmolecule inhibition of PKC suppressed RelA activity, while blocking NF-κB suppressed Twist1 and AR expression. Conversely, inhibition of AR signaling by androgen depletion and the novel antiandrogen enzalutamide induced PKC and RelA activation, resulting in Twist1/AR induction at the transcript level. Moreover, inhibition of NF-κB signaling prevented enzalutamide-induced Twist1 and AR induction. Finally, NF-κB was activated in both castration-resistant and enzalutamide-resistant cells. In conclusion, NF-κB signaling was responsible for Twist1 upregulation by PKC in response to AR inhibition, resulting in aberrant activation of AR. NF-κB signaling thus appears to play a critical role in promoting both castration resistance and enzalutamide resistance in PKC/Twist1 signaling in prostate cancer..
20. Jun Yokoyama, hiroo yamaguchi, Hiroshi Shigeto, Takeshi Uchiumi, Hiroyuki Murai, Jun-Ichi Kira, A case of rhabdomyolysis after status epilepticus without stroke-like episodes in mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes, Clinical Neurology, 10.5692/clinicalneurol.cn-001044, 57, 7, 400-401, 2017.01.
21. Eiichi Ogawa, Norihiro Furusyo, Murata Masayuki, Motohiro Shimizu, Kazuhiro Toyoda, Taeko Hotta, Takeshi Uchiumi, Jun Hayashi, Comparison of the Abbott RealTime HCV and Roche COBAS Ampliprep/COBAS TaqMan HCV assays for the monitoring of sofosbuvir-based therapy, Antiviral Therapy, 10.3851/IMP3085, 22, 1, 61-70, 2017.01, Background: On-treatment HCV kinetics play an invaluable role in evaluating the efficacy of interferon-based therapies. However, the importance of HCV RNA monitoring has not been well discussed concerning treatment with sofosbuvir (SOF)-based regimens, especially for the utility of the Abbott RealTime HCV (ART) assay. Methods: This study consisted of 151 patients infected with HCV genotype-1 or -2, including patients with prior treatment-experience or cirrhosis. HCV genotype-1 patients were treated with SOF/ledipasvir and genotype-2 patients with SOF/ribavirin, both for 12 weeks. Serial measurements of HCV RNA were performed with both the ART and COBAS AmpliPrep/COBAS TaqMan v2.0 (CAP/CTM) assays simultaneously at weeks 0, 1, 2, 4, 6, 8, 10 and 12 of treatment. Results: The rates of HCV RNA target not detected (TND) by ART were significantly lower than those by CAP/CTM between weeks 2 and 12 (end of treatment [EOT]), irrespective of prior treatment-experience or cirrhosis. 11 (11.6%) genotype-1 and 8 (14.3%) genotype-2 patients did not achieve HCV RNA TND by ART at EOT, in contrast to all having HCV RNA TND by CAP/CTM; however, all achieved sustained virological response. The time at which HCV RNA became TND or unquantifiable was not associated with treatment outcome by either the ART or CAP/CTM assay. Conclusions: Over 10% of the patients continued to have detectable HCV RNA by ART at EOT, irrespective of HCV genotype, prior treatment-experience and/or cirrhosis. However, prolonged residual HCV RNA was not associated with treatment failure..
22. masaki shiota, N. Fujimoto, M. Itsumi, ario takeuchi, Junichi Inokuchi, Katsunori Tatsugami, A. Yokomizo, Shunichi Kajioka, Takeshi Uchiumi, Masatoshi Eto, Gene polymorphisms in antioxidant enzymes correlate with the efficacy of androgen-deprivation therapy for prostate cancer with implications of oxidative stress, Annals of Oncology, 10.1093/annonc/mdw646, 28, 3, 569-575, 2017.01, Background: Oxidative stress mitigated by antioxidant enzymes is thought to be involved in the progression to castrationresistant prostate cancer (CRPC) during androgen-deprivation therapy (ADT). This study investigated the association between genetic variations in antioxidant enzymes and the efficacy of ADT as well as its biological background. Patients and methods: The non-synonymous or promoter-locating polymorphisms of antioxidant enzymes were examined as well as the time to CRPC progression and overall survival in 104 and 92 patients treated with ADT for metastatic and nonmetastatic prostate cancer, respectively. In addition, intracellular reactive oxygen species and expression levels of antioxidant enzymes were examined in castration-resistant and enzalutamide-resistant cells. Results: In metastatic prostate cancer, the AG/GG allele in GSTM3 rs7483 and CT/TT allele in CAT rs564250 were associated with a significantly lower risk of progression to CRPC and all-cause death compared with homozygotes of the major AA allele (hazard ratio [HR]; [95% confidence interval (CI)], 0.55 [0.34-0.86], P=0.0086) and CC allele (HR; [95% CI], 0.48 [0.24-0.88], P=0.016), respectively. On multivariate analyses, only GSTM3 rs7483 was associated with significant progression risk (AG/GG versus AA; HR; [95% CI], 0.45 [0.25-0.79], P=0.0047) even after Bonferroni adjustment. In non-metastatic prostate cancer, the AG/GG allele in GSTM3 rs7483 was associated with a significantly lower risk of progression to CRPC (HR; [95% CI], 0.35 [0.10-0.93], P=0.034) and all-cause death (HR; [95% CI], 0.26 [0.041-0.96], P=0.043) compared with the AA allele. Intracellular reactive oxygen species levels were increased, accompanied with augmented GSTM3 expression in both castration-resistant and enzalutamide-resistant cells. Conclusions: Differential activity of antioxidant enzymes caused by the polymorphism in GSTM3 may contribute to resistance to hormonal therapy through oxidative stress. The GSTM3 rs7483 polymorphism may be a promising biomarker for prostate cancer patients treated with ADT..
23. Takeshi Uchiumi, Dongchon Kang, Mitochondrial nucleic acid binding proteins associated with diseases, Frontiers in Bioscience - Landmark, 10.2741/4479, 22, 168-179, 2017.01, Mammalian mitochondrial DNA (mtDNA) exists in structures called nucleoids, which correspond to the configuration of nuclear DNA. Mitochondrial transcription factor A (TFAM), first cloned as an mtDNA transcription factor, is critical for packaging and maintaining mtDNA. To investigate functional aspects of TFAM, we identified many RNA-binding proteins as candidate TFAM interactors, including ERAL1 and p32. In this review, we first describe the functions of TFAM, replication proteins such as polymerase gamma and Twinkle, and mitochondrial RNA binding proteins. We describe the role of mitochondrial nucleic acid binding proteins within the mitochondrial matrix and two oxidative phosphorylation-related proteins within the mitochondrial intermembrane space. We then discuss how mitochondrial dysfunction is related to several diseases, including mitochondrial respiratory disease, Miller syndrome and cancer. We also describe p32 knockout mice, which are embryonic lethal and exhibit respiratory chain defects. Miller syndrome is a recessive disorder characterized by postaxial acrofacial dysostosis and caused by a mutation in DHODH. Finally, we explain that p32 and mitochondrial creatine kinase may be novel markers for the progression of prostate cancer..
24. Hideya Ando, Yoshiaki Ohagi, Moemi Yoshida, Satoshi Yoshimoto, Yusuke Higashi, Masayuki Yagi, Keisuke Monji, Mikako Yagi, Takeshi Uchiumi, Dongchon Kang, Masamitsu Ichihashi, Melanin pigment interrupts the fluorescence staining of mitochondria in melanocytes, Journal of Dermatological Science, 10.1016/j.jdermsci.2016.08.533, 84, 3, 349-351, 2016.12.
25. Amamoto R, Uchiumi T, Yagi M, Monji K, Song Y, Oda Y, , The Expression of Ubiquitous Mitochondrial Creatine Kinase Is Downregulated as Prostate Cancer Progression. , J Cancer , 10.7150/jca.13207, 7, 50-59, 2016.07, BACKGROUND: Mitochondria play crucial roles in cell signaling events, interorganellar communication, aging, cell proliferation and apoptosis, and mitochondrial impairment has been shown to accelerate or modulate cancer progression. Ubiquitous mitochondrial creatine kinase (uMtCK) is predominantly localized in the intermembrane space of mitochondria and catalyzes the reversible exchange of high-energy phosphate between adenosine tri-phosphate (ATP) and phosphocreatine. However, little is known about its expression and function in human prostate cancer progression. METHOD: We investigated the expression of uMtCK in 148 prostate carcinoma tissues and matched normal tissue by immunohistochemistry. The expression and localization of uMtCK and hexokinase II, a marker of glycolysis, were examined in prostate carcinoma cell lines using western blot and immunofluorescence. RESULTS: MtCK expression was significantly lower in high Gleason grade carcinoma compared with normal prostate or low grade carcinoma. Western blot further revealed that uMtCK was highly expressed in LNCaP and 22Rv1 cell lines, as well as in the normal prostate cell line RWPE-1. However, uMtCK expression was almost absent in PC3 and DU145 cell lines, in correlation with absent or mutant p53 expression, respectively. In contrast, hexokinase II was overexpressed in PC3 cells. Moreover, in the low uMtCK expressing cell lines, glycolytic ATP production was increased, whereas mitochondrial ATP production was decreased. CONCLUSIONS: These data suggest that uMtCK is downregulated as prostate cancer progresses in correlation with a metabolic switch in ATP usage..
26. Momoe Itsumi, masaki shiota, ario takeuchi, Eiji Kashiwagi, Junichi Inokuchi, Katsunori Tatsugami, Shunichi Kajioka, Takeshi Uchiumi, Seiji Naito, Masatoshi Eto, Akira Yokomizo, Equol inhibits prostate cancer growth through degradation of androgen receptor by S-phase kinase-associated protein 2, Cancer Science, 10.1111/cas.12948, 107, 7, 1022-1028, 2016.07, Chemopreventive and potential therapeutic effects of soy isoflavones have been shown to be effective in numerous preclinical studies as well as clinical studies in prostate cancer. Although the inhibition of androgen receptor signaling has been supposed as one mechanism underlying their effects, the precise mechanism of androgen receptor inhibition remains unclear. Thus, this study aimed to clarify their mechanism. Among soy isoflavones, equol suppressed androgen receptor as well as prostate-specific antigen expression most potently in androgen-dependent LNCaP cells. However, the inhibitory effect on androgen receptor expression and activity was less prominent in castration-resistant CxR and 22Rv1 cells. Consistently, cell proliferation was suppressed and cellular apoptosis was induced by equol in LNCaP cells, but less so in CxR and 22Rv1 cells. We revealed that the proteasome pathway through S-phase kinase-associated protein 2 (Skp2) was responsible for androgen receptor suppression. Taken together, soy isoflavones, especially equol, appear to be promising as chemopreventive and therapeutic agents for prostate cancer based on the fact that equol augments Skp2-mediated androgen receptor degradation. Moreover, because Skp2 expression was indicated to be crucial for the effect of soy isoflavones, soy isoflavones may be applicable for precancerous and cancerous prostates..
27. Jingxian Fang, Haruyoshi Yamaza, Takeshi Uchiumi, Yoshihiro Hoshino, Keiji Masuda, Yuta Hirofuji, Frank A.D.T.G. Wagener, Dongchon Kang, Kazuaki Nonaka, Dihydroorotate dehydrogenase depletion hampers mitochondrial function and osteogenic differentiation in osteoblasts, European Journal of Oral Sciences, 10.1111/eos.12270, 124, 3, 241-245, 2016.06, Mutation of the dihydroorotate dehydrogenase (DHODH) gene is responsible for Miller syndrome, which is characterized by craniofacial malformations with limb abnormalities. We previously demonstrated that DHODH was involved in forming a mitochondrial supercomplex and that mutated DHODH led to protein instability, loss of enzyme activity, and increased levels of reactive oxygen species in HeLa cells. To explore the etiology of Miller syndrome in more detail, we investigated the effects of DHODH inhibition in the cells involved in skeletal structure. Dihydroorotate dehydrogenase in MC3T3-E1 cells derived from mouse calvaria osteoblast precursor cells was knocked down by specific small interfering RNAs (siRNAs), and cell proliferation, ATP production, and expression of bone-related genes were investigated in these cells. After depletion of DHODH using specific siRNAs, inhibition of cell proliferation and cell cycle arrest occurred in MC3T3-E1 cells. In addition, ATP production was reduced in whole cells, especially in mitochondria. Furthermore, the levels of runt-related transcription factor 2 (Runx2) and osteocalcin (Ocn) mRNAs were lower in DHODH siRNA-treated cells compared with controls. These data suggest that depletion of DHODH affects the differentiation and maturation of osteoblasts. This study shows that mitochondrial dysfunction by DHODH depletion in osteoblasts can be directly linked to the abnormal bone formation in Miller syndrome..
28. Hideyuki Ikematsu, Yong Jeong, Kenjiro Shirane, Hidehiro Toh, Hiroyuki Sasaki, Shinya Matsumoto, Nozomi Noda, Taeko Hotta, Takeshi Uchiumi, Dongchon Kang, Neuraminidase Amino Acid Sequences of Influenza A/H3N2 and B Viruses Isolated from Influenza Patients in the 2014/15 Japanese Influenza Season, Fukuoka Acta Medica, 107, 5, 98-104, 2016.05, Background: Neuraminidase (NA) is a surface protein essential for influenza virus replication. NA inhibitors are commonly used for the treatment of influenza patients in Japan. Several mutations that reduce the effect of NA inhibitors have been reported. We sequenced the whole NA segment of isolated virus from influenza patients and investigated the relation between the NA amino acid sequence and the 50% inhibitory concentration (IC_50) of four NA inhibitors.
Materials and Methods: Forty A/H3N2 and 19 B influenza virus isolated from patients in the 2014/15 influenza season were analyzed. The IC_50 was determined by a neuraminidase inhibition assay using a fluorescent substrate. Viral RNA was amplified by RT-PCR and the genome was sequenced using a next generation sequencer. The deduced amino acid sequences were analyzed.
Results: There was no AA change in the NA catalytic site of the A/H3N2 and B viruses isolated in the 2014-15 influenza season. There was no significant relation between the NA amino acids and the IC_50 of the four NA inhibitors for A/H3N2 or B viruses.
Conclusion: The catalytic site of NA was highly conserved for these A/H3N2 and B viruses. No emergence of NA amino acid mutations related to the sensitivity of the four currently used NA inhibitors was observed..
29. Eiichi Ogawa, Norihiro Furusyo, Murata Masayuki, Takeo Hayashi, Motohiro Shimizu, Haru Mukae, Kazuhiro Toyoda, Taeko Hotta, Takeshi Uchiumi, Jun Hayashi, Impact of HCV kinetics on treatment outcome differs by the type of real-time HCV assay in NS3/4A protease inhibitor-based triple therapy, Antiviral Research, 10.1016/j.antiviral.2015.12.001, 126, 35-42, 2016.02, Repeated measurement of the HCV RNA level is essential for properly monitoring treatment efficacy. The aim of this study was to determine the utility of two HCV real-time assays in the evaluation of the impact of hepatitis C virus (HCV) kinetics on the outcome of triple therapy with NS3/4A protease inhibitors (PIs), telaprevir or simeprevir. This study consisted of 171 Japanese patients infected with HCV genotype 1. All 3266 serum samples taken during and post treatment were tested with both the COBAS AmpliPrep/COBAS TaqMan (CAP/CTM) HCV Test v2.0 and the Abbott RealTime (ART) HCV Test. Of the 2597 samples undetectable (lower limit of detection [
30. Jun Yokoyama, hiroo yamaguchi, Hiroshi Shigeto, Takeshi Uchiumi, Hiroyuki Murai, Jun-Ichi Kira, A case of rhabdomyolysis after status epilepticus without stroke-like episodes in mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes, Clinical Neurology, 10.5692/clinicalneurol.cn-000834, 56, 3, 204-207, 2016.01, A 24-year-old man was referred to our hospital emergency department due to a sudden onset of convulsions after drinking. On arrival he presented status epilepticus and was managed by artificial ventilation. He had no brainstem signs or meningeal irritation. Head MRI showed an old infarction-like lesion in the left occipital lobe, but no abnormal signals on diffusion-weighted images. The patient showed acute rhabdomyolysis (CK 18,000 IU/l) and renal failure, and hemodialysis was started. On 18 day after admission, he was transferred to our department with mild proximal limb muscle weakness and bilateral sensorineural hearing impairment. Electroencephalography demonstrated diffuse intermittent slow wave activities. We suspected a mitochondrial disease because of a significant increase in the lactate/pyruvate ratio (24.1) in the spinal fluid, and identified A3243G mutations in mitochondrial DNA (heteroplasmy 20%) in peripheral white blood cells. We diagnosed his illness as mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). This is a rare case presenting an acute onset of rhabdomyolysis following alcohol intake related to A3243G mitochondrial mutation without preceding stroke-like episodes..
31. Kyousuke Toyoda, Takao Fukuda, Terukazu Sanui, Urara Tanaka, Kensuke Yamamichi, Ryo Atomura, Hidefumi Maeda, Atsushi Tomokiyo, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Grp78 Is Critical for Amelogenin-Induced Cell Migration in a Multipotent Clonal Human Periodontal Ligament Cell Line, Journal of Cellular Physiology, 10.1002/jcp.25087, 231, 2, 414-427, 2016.01, Periodontal ligament stem cells (PDLSCs) are known to play a pivotal role in regenerating the periodontium. Amelogenin, which belongs to a family of extracellular matrix (ECM) proteins, is a potential bioactive molecule for periodontal regenerative therapy. However, its downstream target molecules and/or signaling patterns are still unknown. Our recent proteomic study identified glucose-regulated protein 78 (Grp78) as a new amelogenin-binding protein. In this study, we demonstrate, for the first time, the cellular responses induced by the biological interaction between amelogenin and Grp78 in the human undifferentiated PDL cell line 1-17, which possesses the most typical characteristics of PDLSCs. Confocal co-localization experiments revealed the internalization of recombinant amelogenin (rM180) via binding to cell surface Grp78, and the endocytosis was inhibited by the silencing of Grp78 in 1-17 cells. Microarray analysis indicated that rM180 and Grp78 regulate the expression profiles of cell migration-associated genes in 1-17 cells. Moreover, Grp78 overexpression enhanced rM180-induced cell migration and adhesion without affecting cell proliferation, while silencing of Grp78 diminished these activities. Finally, binding of rM180 to Grp78 promoted the formation of lamellipodia, and the simultaneous activation of Rac1 was also demonstrated by NSC23766, a widely accepted Rac1 inhibitor. These results suggest that Grp78 is essential for enhancing amelogenin-induced migration in 1-17 cells. The biological interaction of amelogenin with Grp78 offers significant therapeutic potential for understanding the biological components and specific functions involved in the signal transduction of amelogenin-induced periodontal tissue regeneration. J. Cell. Physiol. 231: 414-427, 2016..
32. masaki shiota, Naohiro Fujimoto, Kenjiro Imada, Akira Yokomizo, Momoe Itsumi, ario takeuchi, Hidetoshi Kuruma, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Yoshinao Oda, Seiji Naito, Potential role for YB-1 in castration-resistant prostate cancer and resistance to enzalutamide through the androgen receptor V7, Journal of the National Cancer Institute, 10.1093/jnci/djw005, 108, 7, 2016.01, Background: Although androgen deprivation therapy for advanced prostate cancer initially exerts excellent anticancer effects, most prostate cancer treated with androgen deprivation therapy eventually recurs as castration-resistant prostate cancer (CRPC). Although aberrant kinase activation has been proposed as a mechanism of castration resistance, comprehensive kinase profiles in CRPC remain unknown. Therefore, we aimed to elucidate the kinome in CRPC as well as the role of key molecules. Methods: We utilized a kinome array in androgen-dependent LNCaP and castration-resistant CxR cells. The effect of Y-box binding protein-1 (YB-1) on androgen receptor (AR) expression was examined by quantitative polymerase chain reaction and western blot analysis. The association between polymorphisms in the YB-1 gene determined by genotyping and YB-1 expression evaluated by immunohistochemistry in prostate cancer tissues, as well as outcome in metastatic prostate cancer, were investigated by the Cochran-Armitage test and the Cox proportional hazards model, respectively. All statistical tests were two-sided. Results: One hundred fifty-six of 180 kinase phosphorylation sites, including ERK and RSK, were activated in CRPC cells, leading to increased phosphorylation of YB-1, which is a key molecule in the progression to CRPC. YB-1 signaling regulated AR V7 expression, and YB-1 inhibition augmented the anticancer effect of enzalutamide. Moreover, polymorphism (rs12030724) in the YB-1 gene affected YB-1 expression in 93 prostate cancer tissues (YB-1 positive rate; 14.3% in TT, 40.0% in AT, and 52.9% in AA, P = .04) and associated with probability of progression in 104 metastatic prostate cancer case patients (AT/TT vs AA, hazard ratio = 0.49, 95% confidence interval = 0.32 to 0.77, P = .001). Conclusions: YB-1 appears to be a promising target to inhibit the development of castration resistance, even at the AR variant-expressing stage. Polymorphism in the YB-1 gene may be a promising predictive biomarker in hormonal therapy..
33. Keisuke Monji, Takeshi Uchiumi, Saki Hoshizawa, Mikako Yagi, Takashi Matsumoto, Daiki Setoyama, Yuichi Matsushima, Kazuhito Gotou, Rie Amamoto, Dongchon Kang, Serum depletion induced cancer stem cell-like phenotype due to nitric oxide synthesis in oncogenic HRas transformed cells, Oncotarget, 10.18632/oncotarget.12117, 7, 46, 75221-75234, 2016.01, Cancer cells rewire their metabolism and mitochondrial oxidative phosphorylation (OXPHOS) to promote proliferation and maintenance. Cancer cells use multiple adaptive mechanisms in response to a hypo-nutrient environment. However, little is known about how cancer mitochondria are involved in the ability of these cells to adapt to a hypo-nutrient environment. Oncogenic HRas leads to suppression of the mitochondrial oxygen consumption rate (OCR), but oxygen consumption is essential for tumorigenesis. We found that in oncogenic HRas transformed cells, serum depletion reversibly increased the OCR and membrane potential. Serum depletion promoted a cancer stem cell (CSC)-like phenotype, indicated by an increase in CSC markers expression and resistance to anticancer agents. We also found that nitric oxide (NO) synthesis was significantly induced after serum depletion and that NO donors modified the OCR. An NOS inhibitor, SEITU, inhibited the OCR and CSC gene expression. It also reduced anchorage-independent growth by promoting apoptosis. In summary, our data provide new molecular findings that serum depletion induces NO synthesis and promotes mitochondrial OXPHOS, leading to tumor progression and a CSC phenotype. These results suggest that mitochondrial OCR inhibitors can be used as therapy against CSC..
34. Shiota M, Akira Yokomizo, Takeshi Uchiumi, Seiji Naito, Crosstalk between epithelial-mesenchymal transition and castration resistance mediated by Twist1/AR signaling in prostate cancer, ENDOCRINE-RELATED CANCER, 10.1530/ERC-15-0225, 22, 6, 889-900, 2015.12.
35. Shiota M, Akira Yokomizo, Takeshi Uchiumi, Seiji Naito, SRD5A gene polymorphism in Japanese men predicts prognosis of metastatic prostate cancer with androgen-deprivation therapy, EUROPEAN JOURNAL OF CANCER, 10.1016/j.ejca.2015.06.122, 51, 14, 1962-1969, 2015.09.
36. Hideyuki Ikematsu, Yong Jeong, Kenjiro Shirane, Hidehiro Toh, Hiroyuki Sasaki, Yui Koga, Shinya Matsumoto, Taeko Hotta, Takeshi Uchiumi, Dongchon Kang, Analysis of the Neuraminidase Amino Acid Sequences of Influenza A/H1N1pdm09, A/H3N2, and B Viruses Isolated from Influenza Patients in the 2013/14 Japanese Influenza Season, Fukuoka Acta Medica, 106, 8, 231-239, 2015.08, BACKGROUND: Neuraminidase (NA) is an essential surface protein for influenza virus replication. NA inhibitors are commonly used for the treatment of influenza patients in Japan. Several mutations that reduce the effect of NA inhibitors have been reported. We sequenced the whole NA segment of isolated virus from influenza patients and investigated the relation between the NA amino acid sequence and the 50% inhibitory concentration (IC50) of four NA inhibitors.
MATERIALS AND METHODS: A total of 20 viruses that showed high or low IC50 of NA inhibitors were selected from A/H1N1pdm09, A/H3N2, and B isolates from the viruses isolated from patients in the 2013-14 influenza season. Viral RNA was extracted and RT-PCR was done. The amplified genome was sequenced using a next generation sequencer", and the deduced amino acid sequences were analyzed.
RESULTS: Two A/H1N1pdm09 viruses that showed very high IC50 for oseltamivir (150 nM and 130 nM) contained the H275Y mutation. Otherwise, no significant relation was found between the NA amino acids and the IC50 of the four NA inhibitors. There was no significant relation between the NA amino acids and the IC50 of the four NA inhibitors for A/H3N2 viruses. The B viruses that showed a high IC50 for oseltamivir and laninamivir shared some amino acids. The B viruses that showed a high IC50 of zanamivir and peramivir also shared some amino acids. They were different from the shared amino acids found for oseltamivir and laninamivir.
CONCLUSION: The previously reported H275Y mutation that causes oseltamivir resistance was found in the two A/H1N1pdm09 viruses that showed a very high IC50 for oseltamivir. No additional NA amino acid sequences related to the IC50 of the four NA inhibitors was found. The meaning of the shared amino acids among B viruses that showed a high IC50 would be an interesting target for further investigation..
37. Kuniyuki Nakamura, Ago T, Takeshi Uchiumi, Kitazono T, A CADASIL-Like Case with a Novel Noncysteine Mutation of the NOTCH3 Gene and Granular Deposits in the Renal Arterioles, Case Rep Neurol Med, 2015.03.
38. Hideyuki Ikematsu, Yong Jeong, Kenjiro Shirane, Hidehiro Toh, Hiroyuki Sasaki, Yui Koga, Michiyo Urata, Taeko Hotta, Takeshi Uchiumi, Dongchon Kang, Analysis of influenza A/H3N2 neuraminidase genes obtained from influenza patients in the 2011/12 and 2012/13 seasons in Japan, Fukuoka Acta Medica, 106, 1, 16-22, 2015.01, BACKGROUND: Influenza virus has neuraminidase (NA), a surface protein with enzymatic activity that is essential for virus replication. Mutation may affect the effectiveness of NA inhibitors that are used for the treatment of influenza patients. In this study, we determined the NA gene sequences from the clinical isolates of influenza patients to examine the chronological genetic changes and the relation to drug susceptibility.
METHODS: For 96 A/H3N2 virus isolates the 50% inhibitory concentration (IC50) (48 each from the 2011-12 and 12-13 influenza seasons) was measured. RT-PCR was done with extracted viral RNA, followed by nucleotide sequencing.
RESULTS: One putative amino acid mutation, D151N, was found in an NA activity-related cite in five of ninety-six tested isolate. The mutation did not affect the IC50 value. The mutations identified at amino acid positions 387 and 400 were statistically correlated with an increased IC50 value, although the change was less than ten times, suggesting no significant difference in the clinical effectiveness. A small number .of isolates showed mutation in the T and/or B cell epitope region of NA.
CONCLUSION: No mutation that affected the IC50 value or effectiveness of NAIs was detected. Antigenic mutations of NA, which influence the selection of epidemic strains, were not determined. Continuous observation will be necessary to further clarify the genetic features of NA..
39. Aihara, M, Jin, X, Kurihara, Y, Yoshida, Y., Yuichi Matsushima, Takeshi Uchiumi, Dongchon Kang, Kanki, T., Tor and the Sin3-Rpd3 complex regulate expression of the mitophagy receptor protein Atg32 in yeast, 10.1242/jcs.153254 jcs.153254 [pii], 127, 3184-3196, 2014.05.
40. Itsumi, M., Shiota, M., Yokomizo, A, Takeshi Uchiumi, PMA induces androgen receptor downregulation and cellular apoptosis in prostate cancer cells, 10.1530/JME-13-0303 JME-13-0303 [pii], 53, 1, 31-41, 2014.05.
41. Masaki Shiota, Akira Yokomizo, Takeshi Uchiumi, Seiji Naito, Inhibition of RSK/YB-1 signaling enhances the anti-cancer effect of enzalutamide in prostate cancer, 10.1002/pros.22813, 74, 9, 959-969, 2014.04.
42. Momoe Itsumi, masaki shiota, Akira Yokomizo, ario takeuchi, Eiji Kashiwagi, Takashi Dejima, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Seiji Naito, PMA induces androgen receptor downregulation and cellular apoptosis in prostate cancer cells, Journal of Molecular Endocrinology, 10.1530/JME-13-0303, 53, 1, 31-41, 2014.04, Phorbol 12-myristate 13-acetate (PMA) induces cellular apoptosis in prostate cancer cells, the growth of which is governed by androgen/androgen receptor (AR) signaling, but the mechanism by which PMA exerts this effect remains unknown. Therefore, in this study, we investigated the mechanistic action of PMA in prostate cancer cells with regard to AR. We showed that PMA decreased E2F1 as well as AR expression in androgen-dependent prostate cancer LNCaP cells. Furthermore, PMA activated JNK and p53 signaling, resulting in the induction of cellular apoptosis. In LNCaP cells, androgen deprivation and a novel anti-androgen enzalutamide (MDV3100) augmented cellular apoptosis induced by PMA. Moreover, castration-resistant prostate cancer (CRPC) C4-2 cells were more sensitive to PMA compared with LNCaP cells and were sensitized to PMA by enzalutamide. Finally, the expression of PKC, E2F1, and AR was diminished in PMA-resistant cells, indicating that the gain of independence from PKC, E2F1, and AR functions leads to PMA resistance. In conclusion, PMA exerted its anti-cancer effects via the activation of pro-apoptotic JNK/p53 and inhibition of pro-proliferative E2F1/AR in prostate cancer cells including CRPC cells. The therapeutic effects of PMA were augmented by androgen deletion and enzalutamide in androgen-dependent prostate cancer cells, as well as by enzalutamide in castrationresistant cells. Taken together, PMA derivatives may be promising therapeutic agents for treating prostate cancer patients including CRPC patients..
43. masaki shiota, Akira Yokomizo, ario takeuchi, Kenjiro Imada, Eiji Kashiwagi, Yoohyun Song, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Seiji Naito, Inhibition of protein kinase C/twist1 signaling augments anticancer effects of androgen deprivation and enzalutamide in prostate cancer, Clinical Cancer Research, 10.1158/1078-0432.CCR-13-1809, 20, 4, 951-961, 2014.02, Purpose: The progression of prostate cancer to metastatic and castration-resistant disease represents a critical step. We previously showed that the transcription factor Twist1, which promotes epithelial- mesenchymal transition, was involved in castration-resistant progression. Similarly, protein kinase C (PKC) has been implicated in both metastatic progression and castration resistance in prostate cancer. Experimental Design: In this study, we aimed to elucidate the role of PKC/Twist1 signaling in castration resistance, and to apply this information to the development of a novel therapeutic concept using PKC inhibitor Ro31-8220 against prostate cancer using various prostate cancer cell lines. Results: Androgen deprivation and the next-generation antiandrogen enzalutamide induced PKC activation and Twist1 expression, which were reversed by the PKC inhibitor Ro31-8220. Ro31-8220 suppressed cell proliferation in androgen-dependent prostate cancer LNCaP cells, which was augmented by its combination with androgen deprivation or enzalutamide. The favorable anticancer effects of the combination of Ro31-8220 and enzalutamide were also observed in castration-resistant C4-2 and 22Rv1 cells. Furthermore, PKC phosphorylation was elevated in castration-resistant and enzalutamide-resistant cells compared with their parental cells, leading to persistent sensitivity to Ro-31-8220 in castration- and enzalutamide-resistant cells. Conclusions: Taken together, these findings indicate that PKC/Twist1 signaling contributes to castration resistance as well as enzalutamide resistance in prostate cancer, and suggest that therapeutics targeting PKC/ Twist1 signaling, such as PKC inhibitors, represent a promising novel therapeutic strategy for prostate cancer, especially castration-resistant prostate cancer, when combined with enzalutamide..
44. Eiji Kashiwagi, masaki shiota, Akira Yokomizo, Junichi Inokuchi, Takeshi Uchiumi, S. Naito, EP2 signaling mediates suppressive effects of celecoxib on androgen receptor expression and cell proliferation in prostate cancer, Prostate Cancer and Prostatic Diseases, 10.1038/pcan.2013.53, 17, 1, 10-17, 2014.01, Background:Non-steroidal anti-inflammatory drugs inhibit the activity of cyclooxygenases (COXs), and their usage reduces the risks associated with prostate cancer. Celecoxib is a selective COX-2 inhibitor and reported to prevent the progression of prostate cancer. However, the mechanisms involved remain unclear. In this study, we investigated the suppression of prostate cancer growth by celecoxib and elucidated the biological relevance of the inhibited pathway in prostate cancer cell lines.Methods:Western blotting, quantitative real-time PCR and cell proliferation assay were used to resolve the mechanism of celecoxib in prostate cancer cell line PC3, LNCaP and their derivatives.Results:Celecoxib induced apoptosis and downregulated EP2, CREB and androgen receptor (AR). Moreover, EP2 antagonist downregulated CREB as well as COX-2 and AR, resulting in the suppression of cell proliferation. Furthermore, EP2 and CREB knockdown induced AR downregulation, indicating that AR suppression by celecoxib is mediated by EP2/CREB signaling.Conclusions:Celecoxib exerts antitumor activity through EP2 signaling regulating AR and COX-2 expression. Furthermore, in addition to celecoxib, therapeutics targeting EP2 may also be promising against prostate cancers..
45. masaki shiota, Momoe Itsumi, Akira Yokomizo, ario takeuchi, Kenjiro Imada, Eiji Kashiwagi, Junichi Inokuchi, Katsunori Tatsugami, Takeshi Uchiumi, Seiji Naito, Targeting ribosomal S6 kinases/Y-box binding protein-1 signaling improves cellular sensitivity to taxane in prostate cancer, Prostate, 10.1002/pros.22799, 74, 8, 829-838, 2014.01, Background Taxanes are the only cytotoxic chemotherapeutic agents proved to prolong the survival in patients with castration-resistant prostate cancer. However, because of intrinsic and acquired resistances to taxanes, their therapeutical efficiencies are modest, bringing only a few months of survival benefit. Y-box binding protein-1 (YB-1) promotes cancer cell resistance to various anticancer treatments, including taxanes. Here, we aimed to elucidate the mechanism of taxane resistance by YB-1 and examined overcoming resistance by targeting YB-1 signaling. Methods Gene and protein expression levels were evaluated by quantitative real-time polymerase chain reaction and Western blot analysis, respectively. We evaluated the sensitivity of prostate cancer cells to taxanes using cytotoxicity assays. Results Natural taxane paclitaxel from Taxus brevifolia activated the Raf-1/extracellular signal-regulated kinase (ERK) pathway, leading to an activation of ribosomal S6 kinases (RSK)/YB-1 signaling. Activated Raf-1/ERK pathway was blunted by YB-1 knockdown in prostate cancer cells, indicating regulation between Raf-1/ERK signaling and YB-1. In addition, ERK or RSK was activated in taxane-resistant prostate cancer cells, resulting in YB-1 activation. YB-1 knockdown as well as RSK inhibition using RSK-specific siRNA or the small molecule inhibitor SL0101 successfully blocked activation of YB-1, leading to suppression of prostate cancer growth and sensitization to paclitaxel. ConclusionS Taken together, these findings indicate that RSK/YB-1 signaling contributes to taxane resistance, and implicate the therapeutics targeting RSK/YB-1 signaling such as RSK inhibitor as a promising novel therapy against prostate cancer, especially in combination with taxane..
46. Masamune Aihara, Xiulian Jin, Yusuke Kurihara, Yutaka Yoshida, Yuichi Matsushima, Masahide Oku, Yuko Hirota, Tetsu Saigusa, Yoshimasa Aoki, Takeshi Uchiumi, Tadashi Yamamoto, Yasuyoshi Sakai, Dongchon Kang, Tomotake Kanki, Tor and the Sin3-Rpd3 complex regulate expression of the mitophagy receptor protein Atg32 in yeast, Journal of Cell Science, 10.1242/jcs.153254, 127, 14, 3184-3196, 2014.01, When mitophagy is induced in Saccharomyces cerevisiae, the mitochondrial outer membrane protein ScAtg32 interacts with the cytosolic adaptor protein ScAtg11. ScAtg11 then delivers the mitochondria to the pre-autophagosomal structure for autophagic degradation. Despite the importance of ScAtg32 for mitophagy, the expression and functional regulation of ScAtg32 are poorly understood. In this study, we identified and characterized the ScAtg32 homolog in Pichia pastoris (PpAtg32). Interestingly, we found that PpAtg32 was barely expressed before induction of mitophagy and was rapidly expressed after induction of mitophagy by starvation. Additionally, PpAtg32 was phosphorylated when mitophagy was induced. We found that PpAtg32 expression was suppressed by Tor and the downstream PpSin3-PpRpd3 complex. Inhibition of Tor by rapamycin induced PpAtg32 expression, but could neither phosphorylate PpAtg32 nor induce mitophagy. Based on these findings, we conclude that the Tor and PpSin3-PpRpd3 pathway regulates PpAtg32 expression, but not PpAtg32 phosphorylation..
47. Yoo Hyun Song, masaki shiota, Akira Yokomizo, Takeshi Uchiumi, Keijiro Kiyoshima, Kentaro Kuroiwa, Yoshinao Oda, Seiji Naito, Twist1 and Y-box-binding protein-1 are potential prognostic factors in bladder cancer, Urologic Oncology: Seminars and Original Investigations, 10.1016/j.urolonc.2012.11.003, 32, 1, 31.e1-31.e7, 2014.01, Objective: To investigate the expression and possible roles of Twist1 and Y-box-binding protein-1 (YB-1) in bladder cancer tissue. Twist1 belongs to the family of basic helix-loop-helix transcription factors. A functional link between Twist1 and YB-1 has recently been determined to play an important role in bladder cancer cell lines. Materials and methods: Frozen samples from 75 patients with bladder cancer were analyzed by quantitative real-time polymerase chain reaction (PCR). Formalin-fixed and paraffin-embedded tissues from 53 patients with bladder cancer were examined by immunohistochemistry. Results: Twist1 transcript levels were positively correlated with YB-1 transcript levels (coefficient of correlation = 0.42, P<0.001), tumor grade (low grade vs. high grade; P<0.001), invasiveness (non-muscle-invasive bladder cancer vs. muscle invasive bladder cancer; P = 0.0018), and metastasis (meta- vs. meta+; P<0.001). YB-1 transcript level was also correlated with grade (P = 0.029) and invasiveness (P = 0.006). By immunohistochemistry, Twist1 expression was also correlated with YB-1 expression (P<0.001). Further, both Twist1 and YB-1 expression were positively correlated with invasiveness (P = 0.007 and P = 0.002, respectively). Patients with high Twist1 expression and high YB-1 expression had lower overall survival rates, compared with patients with low expression (log-rank test, P = 0.040 and P<0.001, respectively). Conclusions: These results suggest a functional link between Twist1 and YB-1, and they indicate that Twist1 and YB-1 promote bladder cancer progression..
48. 神吉智久, 内海 健, Casein kinase 2 is essential for mitophagy, EMBO Reports, 14, 788-794, 2013.10.
49. Takao Fukuda, Terukazu Sanui, Kyousuke Toyoda, Urara Tanaka, Takaharu Taketomi, Takeshi Uchiumi, Fusanori Nishimura, Identification of Novel Amelogenin-Binding Proteins by Proteomics Analysis, PloS one, 10.1371/journal.pone.0078129, 8, 10, 2013.10, Emdogain (enamel matrix derivative, EMD) is well recognized in periodontology. It is used in periodontal surgery to regenerate cementum, periodontal ligament, and alveolar bone. However, the precise molecular mechanisms underlying periodontal regeneration are still unclear. In this study, we investigated the proteins bound to amelogenin, which are suggested to play a pivotal role in promoting periodontal tissue regeneration. To identify new molecules that interact with amelogenin and are involved in osteoblast activation, we employed coupling affinity chromatography with proteomic analysis in fractionated SaOS-2 osteoblastic cell lysate. In SaOS-2 cells, many of the amelogenin-interacting proteins in the cytoplasm were mainly cytoskeletal proteins and several chaperone molecules of heat shock protein 70 (HSP70) family. On the other hand, the proteomic profiles of amelogenin-interacting proteins in the membrane fraction of the cell extracts were quite different from those of the cytosolic-fraction. They were mainly endoplasmic reticulum (ER)-associated proteins, with lesser quantities of mitochondrial proteins and nucleoprotein. Among the identified amelogenin-interacting proteins, we validated the biological interaction of amelogenin with glucose-regulated protein 78 (Grp78/Bip), which was identified in both cytosolic and membrane-enriched fractions. Confocal co-localization experiment strongly suggested that Grp78/Bip could be an amelogenin receptor candidate. Further biological evaluations were examined by Grp78/Bip knockdown analysis with and without amelogenin. Within the limits of the present study, the interaction of amelogenin with Grp78/Bip contributed to cell proliferation, rather than correlate with the osteogenic differentiation in SaOS-2 cells. Although the biological significance of other interactions are not yet explored, these findings suggest that the differential effects of amelogenin-derived osteoblast activation could be of potential clinical significance for understanding the cellular and molecular bases of amelogenin-induced periodontal tissue regeneration..
50. Tomotake Kanki, Yusuke Kurihara, Xiulian Jin, Tadahiro Goda, Yusuke Ono, Masamune Aihara, Yuko Hirota, Tetsu Saigusa, Yoshimasa Aoki, Takeshi Uchiumi, Dongchon Kang, Casein kinase 2 is essential for mitophagy, EMBO Reports, 10.1038/embor.2013.114, 14, 9, 788-794, 2013.09, Mitophagy is a process that selectively degrades mitochondria. When mitophagy is induced in yeast, the mitochondrial outer membrane protein Atg32 is phosphorylated, interacts with the adaptor protein Atg11 and is recruited into the vacuole with mitochondria. We screened kinase-deleted yeast strains and found that CK2 is essential for Atg32 phosphorylation, Atg32-Atg11 interaction and mitophagy. Inhibition of CK2 specifically blocks mitophagy, but not macroautophagy, pexophagy or the Cvt pathway. In vitro, CK2 phosphorylates Atg32 at serine 114 and serine 119. We conclude that CK2 regulates mitophagy by directly phosphorylating Atg32..
51. masaki shiota, Eiji Kashiwagi, Akira Yokomizo, ario takeuchi, Takashi Dejima, Yoohyun Song, Katsunori Tatsugami, Junichi Inokuchi, Takeshi Uchiumi, Seiji Naito, Interaction between docetaxel resistance and castration resistance in prostate cancer
Implications of Twist1, YB-1, and androgen receptor, Prostate, 10.1002/pros.22681, 73, 12, 1336-1344, 2013.09, BACKGROUND Taxanes, including docetaxel, are currently the only cytotoxic chemotherapeutic agents proven to confer survival benefit in patients with castration-resistant prostate cancer (CRPC). However, the merits of taxanes remain modest, and efforts are needed to improve their therapeutic efficacy. METHODS We evaluated the sensitivity of prostate cancer cells to various agents using cytotoxicity assays. Gene and protein expression levels were evaluated by quantitative real-time polymerase chain reaction and Western blotting analysis, respectively. RESULTS Hydrogen peroxide-resistant and castration-resistant cells that overexpressed Twist1 and Y-box binding protein-1 (YB-1) were cross-resistant to cytotoxic agents, including docetaxel. Twist1 regulated YB-1 expression in prostate cancer cells, supported by the induction of Twist1 and YB-1 by transforming-growth factor-β, which is critical for taxane resistance. Twist1 and/or YB-1 were activated in docetaxel-resistant prostate cancer cells, and YB-1 was activated by docetaxel treatment. Conversely, Twist1 and YB-1 knockdown sensitized prostate cancer cells to cytotoxic agents, including docetaxel. In addition, androgen receptor (AR) knockdown increased cellular sensitivity to docetaxel, though AR expression in docetaxel-resistant LNCaP cells was paradoxically lower than in parental cells. Intriguingly, androgen deprivation treatment was more effective in docetaxel-resistant LNCaP cells compared with parental cells. CONCLUSIONS Twist1/YB-1 and AR signaling promote docetaxel resistance in CRPC cells. However, docetaxel-resistant cells were collaterally sensitive to androgen deprivation because of down-regulation of AR expression, suggesting that the therapeutic effect of initial taxane treatment in hormone-naïve prostate cancer may be superior to that of salvage taxane treatment in CRPC..
52. Eiji Kashiwagi, masaki shiota, Akira Yokomizo, Momoe Itsumi, Junichi Inokuchi, Takeshi Uchiumi, Seiji Naito, Prostaglandin receptor EP3 mediates growth inhibitory effect of aspirin through androgen receptor and contributes to castration resistance in prostate cancer cells, Endocrine-Related Cancer, 10.1530/ERC-12-0344, 20, 3, 431-441, 2013.06, Although numerous epidemiological studies show aspirin to reduce risk of prostate cancer, the mechanism of this effect is unclear. Here, we first confirmed that aspirin downregulated androgen receptor (AR) and prostate-specific antigen in prostate cancer cells. We also found that aspirin upregulated prostaglandin receptor subtype EP3 but not EP2 or EP4. The EP3 antagonist L798106 and EP3 knockdown increased AR expression and cell proliferation, whereas the EP3 agonist sulprostone decreased them, indicating that EP3 affects AR expression. Additionally, EP3 (PTGER3) transcript levels were significantly decreased in human prostate cancer tissues compared with those in normal human prostate tissues, suggesting that EP3 is important to prostate carcinogenesis. Decreased EP3 expression was also seen in castration-resistant subtype CxR cells compared with parental LNCaP cells. Finally, we found that aspirin and EP3 modulators affected prostate cancer cell growth. Taken together, aspirin suppressed LNCaP cell proliferation via EP3 signaling activation; EP3 downregulation contributed to prostate carcinogenesis and to progression from androgen-dependent prostate cancer to castration-resistant prostate cancer by regulating AR expression. In conclusion, cyclooxygenases and EP3 may represent attractive therapeutic molecular targets in androgen-dependent prostate cancer..
53. Jing Xian Fang, Takeshi Uchiumi, Mikako Yagi, Shinya Matsumoto, Rie Amamoto, Shinya Takazaki, Haruyoshi Yamaza, Kazuaki Nonaka, Dongchon Kang, Dihydro-orotate dehydrogenase is physically associated with the respiratory complex and its loss leads to mitochondrial dysfunction, Bioscience Reports, 10.1042/BSR20120097, 33, 2, 217-227, 2013.05, Some mutations of the DHODH (dihydro-orotate dehydrogenase) gene lead to postaxial acrofacial dysostosis or Miller syndrome. Only DHODH is localized at mitochondria among enzymes of the de novo pyrimidine biosynthesis pathway. Since the pyrimidine biosynthesis pathway is coupled to the mitochondrial RC (respiratory chain) via DHODH, impairment of DHODH should affect the RC function. To investigate this, we used siRNA (small interfering RNA)-mediated knockdown and observed that DHODH knockdown induced cell growth retardation because of G2/M cell-cycle arrest, whereas pyrimidine deficiency usually causes G1/S arrest. Inconsistent with this, the cell retardation was not rescued by exogenous uridine, which should bypass the DHODH reaction for pyrimidine synthesis. DHODH depletion partially inhibited the RC complex III, decreased the mitochondrial membrane potential, and increased the generation of ROS (reactive oxygen species). We observed that DHODH physically interacts with respiratory complexes II and III by IP (immunoprecipitation) and BN (blue native)/SDS/PAGE analysis. Considering that pyrimidine deficiency alone does not induce craniofacial dysmorphism, the DHODH mutations may contribute to the Miller syndrome in part through somehow altered mitochondrial function..
54. Takeshi Uchiumi, Hiroyuki Tanamachi, Kajiyo Kuchiwaki, Mitsuharu Kajita, Shinya Matsumoto, Mikako Yagi, Tomotake Kanki, Dongchon Kang, Mutation and functional analysis of ABCC2/multidrug resistance protein 2 in a Japanese patient with Dubin-Johnson syndrome, Hepatology Research, 10.1111/j.1872-034X.2012.01103.x, 43, 5, 569-575, 2013.05, Dubin-Johnson syndrome (DJS) is a recessive inherited disorder characterized by conjugated hyperbilirubinemia. It is caused by dysfunction of adenosine triphosphate-binding cassette, sub-family C, member 2 (ABCC2/MRP2) on the canalicular membrane of hepatocytes. We performed mutational analysis of the ABCC2/MRP2 gene in a Japanese female with DJS. Furthermore, we investigated the effects of the two identified DJS-associated mutations on MRP2 function. We found a compound heterozygous mutation in the patient: W709R (c.2124T>C), a missense mutation in exon 17, and R1310X (c.3928C>T), a nonsense mutation in exon 28. DJS-associated mutations have been shown to impair the protein maturation and transport activity of ABCC2/MRP2. We established HEK293 cell lines stably expressing one of the two identified DJS-associated mutations. Expressed W709R MRP2 was mainly core-glycosylated, predominantly retained in the endoplasmic reticulum, and exhibited no transport activity, suggesting that this mutation causes deficient maturation and impaired protein sorting. No MRP2 protein was expressed from HEK293 cells transfected with an R1310X-containing construct. This compound heterozygous mutation of the MRP2 gene causes dysfunction of the MRP2 protein and the hyperbilirubinemia seen in DJS..
55. Yui Koga, Michiyo Urata, Takeshi Uchiumi, Osamu Sato, Hiroaki Kobayashi, Dongchon Kang, The clinical evaluation of high risk human papillomavirus (HPV) genotyping in cervical cancer tissue using a novel electric DNA chip-based diagnostic system, Japanese Journal of Clinical Chemistry, 42, 2, 140-145, 2013.04, Continuous infection of human papillomavirus (HPV) is known to be the major cause of cervical carcinoma. So far, more than 100 different HPV genotypes have been identified, and more than 13 kinds of HPV are known as high risk types of cervical cancer. Recently, an electric DNA chip-based diagnostic system which detects 13 HPV genotypes easily and quickly was developed. Using the electric DNA chip-based diagnostic system we performed the high risk HPV genotyping in 60 cervical cancer tissue sections and identified HPV DNA in 52 samples (86.7%). This result is consistent with the previous reports that detected the 13 high risk types of HPV from more than 90% of cervical tumor tissues. Among the HPV positive samples, HPV-16 was the most (71.2%[37cases/52cases]) and HPV-18 was the second (30.8%[16/52]) common types, followed by HPV-58(13.5%[7/52]) and -52(9.6%[5/52]). HPV-58 and -52 are prevalent HPV types in Asia and our result is in agreement with this information. There were also eight HPV-negative samples in this study, including one case with minimal deviation adenocarcinoma that is considered to develop without HPV infection. Although the HPV vaccines, such as bivalent and quadrivalent vaccines are commonly used in these days, we observed 16 cases which may not be protected by these vaccines, suggesting that these vaccines do not always protect HPV-induced carcinogenesis..
56. Fang J, 内海 健, 康東天, Dihydroorotate dehydrogenase is physically associated with the respiratory complex and its loss leads to mitochondrial dysfunction., Bioscience report, 33, 217-227, 2013.03.
57. Momoe Itsumi, masaki shiota, Akira Yokomizo, Eiji Kashiwagi, ario takeuchi, Katsunori Tatsugami, Junichi Inokuchi, Yoohyun Song, Takeshi Uchiumi, Seiji Naito, Human heterochromatin protein 1 Isoforms regulate androgen receptor signaling in prostate cancer, Journal of Molecular Endocrinology, 10.1530/JME-13-0024, 50, 3, 401-409, 2013.03, Androgen receptor (AR) signaling is critical for the tumorigenesis and development of prostate cancer, as well as the progression to castration-resistant prostate cancer. We previously showed that the heterochromatin protein 1 (HP1) b isoform plays a critical role in transactivation of AR signaling as an AR coactivator that promotes prostate cancer cell proliferation. However, the roles of other HP1 isoforms, HP1a and HP1g, in AR expression and prostate cancer remain unclear. Here, we found that knockdown of HP1g, but not HP1a, reduced AR expression and cell proliferation by inducing cell cycle arrest at G1 phase in LNCaP cells. Conversely, overexpression of full-length HP1a and its C-terminal deletion mutant increased AR expression and cell growth, whereas overexpression of HP1g had no effect. Similarly, HP1a overexpression promoted 22Rv1 cell growth, whereas HP1g knockdown reduced the proliferation of CxR cells, a castration-resistant LNCaP derivative. Taken together, HP1 isoforms distinctly augment AR signaling and cell growth in prostate cancer. Therefore, silencing of HP1b and HP1g may be a promising therapeutic strategy for treatment of prostate cancer..
58. 内海 健, Mutation and functional analysis of ABCC2/multidrug resistance protein 2 in a Japanese patient with Dubin-Johnson syndrome., Hepatol. Research, 43, 569-575, 2013.02.
59. Takeshi Uchiumi, Dongchon Kang, [The functional and pathological analysis of mitochondrial protein p32]., Unknown Journal, 61, 6, 493-500, 2013.01, p32 is an evolutionarily conserved and ubiquitously expressed multifunctional protein. Although p32 exists at diverse intra and extracellular sites, it is predominantly localized to the mitochondrial matrix near the nucleoid associated with mitochondrial transcription factor A. p32-deficient mice exhibited mid-gestation lethality associated with a severe developmental defect of the embryo. Primary embryonic fibroblasts isolated from p32-knockout embryos showed severe dysfunction of the mitochondrial respiratory chain because of severely impaired mitochondrial protein synthesis. The RNA-binding ability of p32 is well correlated with mitochondrial translation. We also found that p32 is highly expressed in prostate tumor samples and its expression is significantly associated with the Gleason score, pathologic stage, and relapse. These data suggest that p32 is critical for prostate cancer cell proliferation and may be a novel marker of clinical progression in prostate cancer..
60. Jing Xian Fang, Takeshi Uchiumi, Mikako Yagi, Shinya Matsumoto, Rie Amamoto, Shinya Takazaki, Haruyoshi Yamaza, Kazuaki Nonaka, Dongchon Kang, Dihydro-orotate dehydrogenase is physically associated with the respiratory complex and its loss leads to mitochondrial dysfunction., Bioscience Reports, 33, 2, 2013, Some mutations of the DHODH (dihydro-orotate dehydrogenase) gene lead to postaxial acrofacial dysostosis or Miller syndrome. Only DHODH is localized at mitochondria among enzymes of the de novo pyrimidine biosynthesis pathway. Since the pyrimidine biosynthesis pathway is coupled to the mitochondrial RC (respiratory chain) via DHODH, impairment of DHODH should affect the RC function. To investigate this, we used siRNA (small interfering RNA)-mediated knockdown and observed that DHODH knockdown induced cell growth retardation because of G2/M cell-cycle arrest, whereas pyrimidine deficiency usually causes G1/S arrest. Inconsistent with this, the cell retardation was not rescued by exogenous uridine, which should bypass the DHODH reaction for pyrimidine synthesis. DHODH depletion partially inhibited the RC complex III, decreased the mitochondrial membrane potential, and increased the generation of ROS (reactive oxygen species). We observed that DHODH physically interacts with respiratory complexes II and III by IP (immunoprecipitation) and BN (blue native)/SDS/PAGE analysis. Considering that pyrimidine deficiency alone does not induce craniofacial dysmorphism, the DHODH mutations may contribute to the Miller syndrome in part through somehow altered mitochondrial function..
61. Jing Xian Fang, Takeshi Uchiumi, Mikako Yagi, Shinya Matsumoto, Rie Amamoto, Toshiro Saito, Shinya Takazaki, Tomotake Kanki, Haruyoshi Yamaza, Kazuaki Nonaka, Dongchon Kang, Protein instability and functional defects caused by mutations of dihydro-orotate dehydrogenase in Miller syndrome patients, Bioscience Reports, 10.1042/BSR20120046, 32, 6, 631-639, 2012.12, Miller syndrome is a recessive inherited disorder characterized by postaxial acrofacial dysostosis. It is caused by dysfunction of the DHODH (dihydroorotate dehydrogenase) gene, which encodes a key enzyme in the pyrimidine de novo biosynthesis pathway and is localized at mitochondria intermembrane space. We investigated the consequence of three missense mutations, G202A, R346W and R135C of DHODH, which were previously identified in patients with Miller syndrome. First, we established HeLa cell lines stably expressing DHODH with Miller syndrome-causative mutations: G202A, R346W and R135C. These three mutant proteins retained the proper mitochondrial localization based on immunohistochemistry and mitochondrial subfractionation studies. The G202A, R346W DHODH proteins showed reduced protein stability. On the other hand, the third one R135C, in which the mutation lies at the ubiquinone-binding site, was stable but possessed no enzymatic activity. In conclusion, the G202A and R346W mutation causes deficient protein stability, and the R135C mutation does not affect stability but impairs the substrate-induced enzymatic activity, suggesting that impairment of DHODH activity is linked to the Miller syndrome phenotype..
62. Mikako Yagi, 内海 健, 康 東天, P32/gC1qR is indispensable for fetal development and mitochondrial translation: Importance of its RNA-binding ability, Nucleic Acid Reserch, 40, 9717--9737., 2012.10.
63. Mikako Yagi, Takeshi Uchiumi, Shinya Takazaki, Bungo Okuno, Masatoshi Nomura, Shin Ichi Yoshida, Tomotake Kanki, Dongchon Kang, P32/gC1qR is indispensable for fetal development and mitochondrial translation
Importance of its RNA-binding ability, Nucleic Acids Research, 10.1093/nar/gks774, 40, 19, 9717-9737, 2012.10, p32 is an evolutionarily conserved and ubiquitously expressed multifunctional protein. Although p32 exists at diverse intra and extracellular sites, it is predominantly localized to the mitochondrial matrix near the nucleoid associated with mitochondrial transcription factor A. Nonetheless, its function in the matrix is poorly understood. Here, we determined p32 function via generation of p32-knockout mice. p32-deficient mice exhibited midgestation lethality associated with a severe developmental defect of the embryo. Primary embryonic fibroblasts isolated from p32-knockout embryos showed severe dysfunction of the mitochondrial respiratory chain, because of severely impaired mitochondrial protein synthesis. Recombinant p32 binds RNA, not DNA, and endogenous p32 interacts with all mitochondrial messenger RNA species in vivo. The RNA-binding ability of p32 is well correlated with the mitochondrial translation. Coimmunoprecipitation revealed the close association of p32 with the mitoribosome. We propose that p32 is required for functional mitoribosome formation to synthesize proteins within mitochondria..
64. Matsumoto S, Uchiumi T, Tanamachi H, Saito T, Yagi M, Takazaki S, Kanki T, Kang D., Ribonucleoprotein Y-box binding protein-1 regulates mitochondrial oxidative phosphorylation (OXPHOS) protein expression after serum stimulation through binding to OXPHOS mRNA, Biochem J, 2012.05.
65. Eiji Kashiwagi, masaki shiota, Akira Yokomizo, Momoe Itsumi, Junichi Inokuchi, Takeshi Uchiumi, Seiji Naito, Downregulation of phosphodiesterase 4B (PDE4B) activates protein kinase A and contributes to the progression of prostate cancer, Prostate, 10.1002/pros.21478, 72, 7, 741-751, 2012.05, Background Prostate cancer is the most commonly diagnosed non-cutaneous cancer in American men. Unfortunately, few successful therapies for castration-resistant prostate cancer (CRPC) exist. The protein kinase A (PKA) pathway is a critical mediator of cellular proliferation and differentiation in various normal and cancerous cells. However, the PKA activity and the mechanism of regulation in CRPC remain unclear. Then, in this study, we intended to reveal the PKA activity and the mechanism of regulation in CRPC. Methods Western blotting, quantitative real-time polymerase chain reaction, cytotoxicity analysis, and cell proliferation assay were used to resolve the regulatory role of PKA in prostate cancer cell line, LNCaP and their derivatives. Results cAMP-specific phosphodiesterase 4B (PDE4B) was downregulated and the PKA pathway was activated in castration-resistant LNCaP derivatives (CxR cells). Rolipram activated the PKA pathway via inhibition of PDE4B, resulting in AR transactivation while the PKA inhibitor, H89 reduced AR transactivation. In response to hydrogen peroxide and in hydrogen peroxide-resistant LNCaP derivatives (HPR50 cells) PDE4B was decreased and as a result PKA activity was increased. Moreover, PDE4B expression was reduced in advanced prostate cancer and PDE4B knockdown promoted castration-resistant growth of LNCaP cells. Conclusions Oxidative stress may suppress PDE4B expression and activate the PKA pathway. The PDE4B/PKA pathway contributed to progression of androgen-dependent prostate cancer to CRPC. This pathway may represent an attractive therapeutic molecular target..
66. Shinya Matsumoto, Takeshi Uchiumi, Toshiro Saito, Mikako Yagi, Shinya Takazaki, Tomotake Kanki, Dongchon Kang, Localization of mRNAs encoding human mitochondrial oxidative phosphorylation proteins, Mitochondrion, 10.1016/j.mito.2012.02.004, 12, 3, 391-398, 2012.05, The mitochondrial oxidative phosphorylation (OXPHOS) proteins are encoded by both nuclear and mitochondrial DNA. The nuclear-encoded OXPHOS mRNAs have specific subcellular localizations, but little is known about which localize near mitochondria. Here, we compared mRNAs in mitochondria-bound polysome fractions with those in cytosolic, free polysome fractions. mRNAs encoding hydrophobic OXPHOS proteins, which insert into the inner membrane, were localized near mitochondria. Conversely, OXPHOS gene which mRNAs were predominantly localized in cytosol had less than one transmembrane domain. The RNA-binding protein Y-box binding protein-1 is localized at the mitochondrial outer membrane and bound to the OXPHOS mRNAs. Our findings offer new insight into mitochondrial co-translational import in human cells..
67. Takeshi Uchiumi, Dongchon Kang, The role of TFAM-associated proteins in mitochondrial RNA metabolism, Biochimica et Biophysica Acta - General Subjects, 10.1016/j.bbagen.2011.08.014, 1820, 5, 565-570, 2012.05, Background: Mammalian mitochondrial DNA (mtDNA) takes on a higher structure called the nucleoid or mitochromosome, which corresponds to that of nuclear DNA. Mitochondrial transcription factor A (TFAM), which was cloned as a transcription factor for mitochondrial DNA, is critical for forming this higher structure and for maintenance of mtDNA. Scope of review: To investigate the functional aspects of the nucleoid, we have identified many RNA-binding proteins to be candidate TFAM interactors, including ERAL1 and p32. Major conclusions: In this review, we would like to describe that ERAL1 binds to the mitochondrial rRNA component of the ribosomal small subunit and is an important constituent of this subunit. p32, which is involved in mitochondrial translation, may be a novel marker of clinical progression in prostate cancer. Here we describe these proteins, all of which are involved in translation within the mitochondrial matrix. General significance: This review highlights the results from the mitochondrial nucleoid research in organic biochemistry. This article is part of a Special Issue entitled Biochemistry of Mitochondria..
68. Shinya Matsumoto, Takeshi Uchiumi, Hiroyuki Tanamachi, Toshiro Saito, Mikako Yagi, Shinya Takazaki, Tomotake Kanki, Dongchon Kang, Ribonucleoprotein Y-box-binding protein-1 regulates mitochondrial oxidative phosphorylation (OXPHOS) protein expression after serum stimulation through binding to OXPHOS mRNA, Biochemical Journal, 10.1042/BJ20111728, 443, 2, 573-584, 2012.04, Mitochondria play key roles in essential cellular functions, such as energy production, metabolic pathways and aging. Growth factor-mediated expression of the mitochondrial OXPHOS (oxidative phosphorylation) complex proteins has been proposed to play a fundamental role in metabolic homoeostasis. Although protein translation is affected by general RNA-binding proteins, very little is known about the mechanism involved in mitochondrial OXPHOS protein translation. In the present study, serum stimulation induced nuclear-encoded OXPHOS protein expression, such as NDUFA9 [NADH dehydrogenase (ubiquinone) 1α subcomplex, 9, 39 kDa], NDUFB8 [NADH dehydrogenase (ubiquinone) 1β subcomplex, 8, 19 kDa], SDHB [succinate dehydrogenase complex, subunit B, iron sulfur (Ip)] and UQCRFS1 (ubiquinol-cytochrome c reductase, Rieske iron-sulfur polypeptide 1), and mitochondrial ATP production, in a translation-dependent manner. We also observed that the major ribonucleoprotein YB-1 (Y-box-binding protein-1) preferentially bound to these OXPHOS mRNAs and regulated the recruitment of mRNAs from inactivemRNPs (messenger ribonucleoprotein particles) to active polysomes. YB-1 depletion led to up-regulation of mitochondrial function through induction of OXPHOS protein translation from inactive mRNP release. In contrast, YB-1 overexpression suppressed the translation of these OXPHOS mRNAs through reduced polysome formation, suggesting that YB-1 regulated the translation of mitochondrial OXPHOS mRNAs through mRNA binding. Taken together, our findings suggest that YB-1 is a critical factor for translation that may control OXPHOS activity..
69. Matsumoto S, Uchiumi T, Saito T, Yagi M, Takazaki S, Kanki T, Kang D, Localization of mRNAs encoding human mitochondrial oxidative phosphorylation proteins., Mitochondrion, 12, 391-398, 2012.03.
70. masaki shiota, Yoohyun Song, ario takeuchi, Akira Yokomizo, Eiji Kashiwagi, Kentaro Kuroiwa, Katsunori Tatsugami, Takeshi Uchiumi, Yoshinao Oda, Seiji Naito, Antioxidant therapy alleviates oxidative stress by androgen deprivation and prevents conversion from androgen dependent to castration resistant prostate cancer, Journal of Urology, 10.1016/j.juro.2011.09.147, 187, 2, 707-714, 2012.02, Prostate cancer progression from androgen dependence to castration resistance results at least in part from oxidative stress induced by androgen deprivation therapy. We elucidated the state and the role of oxidative stress induced by androgen deprivation therapy and the possibility of antioxidant therapy in human prostate cancer. We investigated 4-HNE (4-hydroxy-2-nonenal histidine adduct) staining, and Twist1, YB-1 and androgen receptor expression by immunohistochemistry in prostate cancer samples treated with or without neoadjuvant androgen deprivation therapy. Intracellular reactive oxygen species and protein expression were examined by CM-H 2DCFDA and Western blot analysis, respectively. A cell proliferation assay and a mouse xenograft model were used to assess tumor growth. Androgen deprivation therapy increased oxidative stress, as shown by 4-HNE staining in human prostate cancer tissue. Twist1 and YB-1 expression was up-regulated by androgen deprivation, resulting in androgen receptor over expression. In LNCaP and 22Rv1 cells androgen deprivation increased intracellular reactive oxygen species and evoked Twist1, YB-1 and androgen receptor over expression, resulting in cell growth in a castration resistant manner. Growth was alleviated by N-acetyl-cysteine, an electrophile that supports glutathione production. N-acetyl-cysteine also decreased LNCaP and 22Rv1 tumor growth in castrated and noncastrated mice. Androgen deprivation therapy induced oxidative stress in in vitro and human prostate cancer. Antioxidant therapy using N-acetyl-cysteine appears to be a promising therapeutic modality for prostate cancer..
71. Yusuke Kurihara, Tomotake Kanki, Yoshimasa Aoki, Yuko Hirota, Tetsu Saigusa, Takeshi Uchiumi, Dongchon Kang, Mitophagy plays an essential role in reducing mitochondrial production of reactive oxygen species and mutation of mitochondrial DNA by maintaining mitochondrial quantity and quality in yeast, Journal of Biological Chemistry, 10.1074/jbc.M111.280156, 287, 5, 3265-3272, 2012.01, In mammalian cells, the autophagy-dependent degradation of mitochondria (mitophagy) is thought to maintain mitochondrial quality by eliminating damaged mitochondria. However, the physiological importance of mitophagy has not been clarified in yeast. Here, we investigated the physiological role of mitophagy in yeast using mitophagy-deficient atg32- or atg11-knock-out cells. When wild-type yeast cells in respiratory growth encounter nitrogen starvation, mitophagy is initiated, excess mitochondria are degraded, and reactive oxygen species (ROS) production from mitochondria is suppressed; as a result, the mitochondria escape oxidative damage. On the other hand, in nitrogen-starved mitophagy-deficient yeast, excess mitochondria are not degraded and the undegraded mitochondria spontaneously age and produce surplus ROS. The surplus ROS damage the mitochondria themselves and the damaged mitochondria produce more ROS in a vicious circle, ultimately leading to mitochondrial DNA deletion and the so-called "petite-mutant"phenotype. Cells strictly regulate mitochondrial quantity and quality because mitochondria produce both necessary energy and harmful ROS. Mitophagy contributes to this process by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production..
72. Yoshimasa Aoki, Tomotake Kanki, Yuko Hirota, Yusuke Kurihara, Tetsu Saigusa, Takeshi Uchiumi, Dongchon Kang, Phosphorylation of serine 114 on Atg32 mediates mitophagy, Molecular biology of the cell, 10.1091/mbc.E11-02-0145, 22, 17, 3206-3217, 2011.09, Mitophagy, which selectively degrades mitochondria via autophagy, has a significant role in mitochondrial quality control. When mitophagy is induced in yeast, mitochondrial residential protein Atg32 binds Atg11, an adaptor protein for selective types of autophagy, and it is recruited into the vacuole along with mitochondria. The Atg11-Atg32 interaction is believed to be the initial molecular step in which the autophagic machinery recognizes mitochondria as a cargo, although how this interaction is mediated is poorly understood. Therefore, we studied the Atg11-Atg32 interaction in detail. We found that the C-terminus region of Atg11, which included the fourth coiled-coil domain, interacted with the N-terminus region of Atg32 (residues 100-120). When mitophagy was induced, Ser-114 and Ser-119 on Atg32 were phosphorylated, and then the phosphorylation of Atg32, especially phosphorylation of Ser-114 on Atg32, mediated the Atg11-Atg32 interaction and mitophagy. These findings suggest that cells can regulate the amount of mitochondria, or select specific mitochondria (damaged or aged) that are degraded by mitophagy, by controlling the activity and/or localization of the kinase that phosphorylates Atg32. We also found that Hog1 and Pbs2, which are involved in the osmoregulatory signal transduction cascade, are related to Atg32 phosphorylation and mitophagy..
73. Fotovati A, Abu-Ali S, Wang PS, Deleyrolle LP, Lee C, Triscott J, Chen JY, Franciosi S, Nakamura Y, Sugita Y, Uchiumi T, Kuwano M, Leavitt BR, Singh SK, Jury A, Jones C, Wakimoto H, Reynolds BA, Pallen CJ, Dunn SE., YB-1 bridges neural stem cells and brain tumor-initiating cells via its roles in differentiation and cell growth., Cancer Res, 71, 5569-78, 2011.08.
74. Aoki Y, Kanki T, Hirota Y, Kurihara Y, Saigusa T, Uchiumi T, Kang D, Phosphorylation of Serine 114 on Atg32 mediates mitophagy., Mol Biol Cell, 22, 3206-17, 2011.08.
75. Uchiumi T, Kang D., The role of TFAM-associated proteins in mitochondrial RNA metabolism., Biochim Biophys Acta., 2011.08.
76. masaki shiota, ario takeuchi, YooHyun Song, Akira Yokomizo, Eiji Kashiwagi, Takeshi Uchiumi, Kentaro Kuroiwa, Katsunori Tatsugami, Naohiro Fujimoto, Yoshinao Oda, Seiji Naito, Y-box binding protein-1 promotes castration-resistant prostate cancer growth via androgen receptor expression, Endocrine-Related Cancer, 10.1530/ERC-11-0017, 18, 4, 505-517, 2011.08, The androgen receptor (AR) is well known to play a central role in the pathogenesis of prostate cancer (PCa). In several studies, AR was overexpressed in castration-resistant PCa (CRPC). However, the mechanism of AR overexpression in CRPC is not fully elucidated. Y-box binding protein-1 (YB-1) is a pleiotropic transcription factor that is upregulated in CPRC. We aimed to elucidate the role of YB-1 in castration resistance of PCa and identify therapeutic potential of targeting YB-1. Using immunohistochemistry, we found that nuclear YB-1 expression significantly correlated with the Gleason score and AR expression in PCa tissues. In PCa cells, YB-1 regulated AR expression at the transcriptional level. Furthermore, YB-1 expression and nuclear localization were upregulated in CRPC cells. Overexpression of AR, as well as YB-1, conferred castrationresistant growth in LNCaP and 22Rv1 cells. Conversely, knocking down YB-1 resulted in suppressed cell growth and induced apoptosis, which was more efficient than knocking down AR in LNCaP cells. In other types of PCa cells, such as CRPC cells, knocking down YB-1 resulted in a significant reduction of cell growth. In conclusion, these findings suggested that YB-1 induces castration resistance in androgen-dependent PCa cells via AR expression. Thus, YB-1 may be a promising therapeutic target for PCa, as well as CRPC..
77. Abbas Fotovati, Samah Abu-Ali, Pei Shan Wang, Loic P. Deleyrolle, Cathy Lee, Joanna Triscott, James Y. Chen, Sonia Franciosi, Yasuhiro Nakamura, Yasuo Sugita, Takeshi Uchiumi, Michihiko Kuwano, Blair R. Leavitt, Sheila K. Singh, Alexa Jury, Chris Jones, Hiroaki Wakimoto, Brent A. Reynolds, Catherine J. Pallen, Sandra E. Dunn, YB-1 bridges neural stem cells and brain tumor-initiating cells via its roles in differentiation and cell growth, Cancer Research, 10.1158/0008-5472.CAN-10-2805, 71, 16, 5569-5578, 2011.08, The Y-box binding protein 1 (YB-1) is upregulated in many human malignancies including glioblastoma (GBM). It is also essential for normal brain development, suggesting that YB-1 is part of a neural stem cell (NSC) network. Here, we show that YB-1 was highly expressed in the subventricular zone (SVZ) of mouse fetal brain tissues but not in terminally differentiated primary astrocytes. Conversely, YB-1 knockout mice had reduced Sox-2, nestin, and musashi-1 expression in the SVZ. Although primary murine neurospheres were rich in YB-1, its expression was lost during glial differentiation. Glial tumors often express NSC markers and tend to loose the cellular control that governs differentiation; therefore, we addressed whether YB-1 served a similar role in cancer cells. YB-1, Sox-2, musashi-1, Bmi-1, and nestin are coordinately expressed in SF188 cells and 9/9 GBM patient-derived primary brain tumor-initiating cells (BTIC). Silencing YB-1 with siRNA attenuated the expression of these NSC markers, reduced neurosphere growth, and triggered differentiation via coordinate loss of GSK3-β. Furthermore, differentiation of BTIC with 1% serum or bone morphogenetic protein-4 suppressed YB-1 protein expression. Likewise, YB-1 expression was lost during differentiation of normal human NSCs. Consistent with these observations, YB-1 expression increased with tumor grade (n = 49 cases). YB-1 was also coexpressed with Bmi-1 (Spearmans 0.80, P > 0.001) and Sox-2 (Spearmans 0.66, P > 0.001) based on the analysis of 282 cases of high-grade gliomas. These proteins were highly expressed in 10/15 (67%) of GBM patients that subsequently relapsed. In conclusion, YB-1 correlatively expresses with NSC markers where it functions to promote cell growth and inhibit differentiation..
78. masaki shiota, Akira Yokomizo, Eiji Kashiwagi, ario takeuchi, Naohiro Fujimoto, Takeshi Uchiumi, Seiji Naito, Peroxiredoxin 2 in the nucleus and cytoplasm distinctly regulates androgen receptor activity in prostate cancer cells, Free Radical Biology and Medicine, 10.1016/j.freeradbiomed.2011.04.001, 51, 1, 78-87, 2011.07, Currently, few therapies are effective against castration-resistant prostate cancer. Increased activation of the androgen/androgen receptor (AR) signaling pathway is thought to promote castration-resistant prostate cancer. Herein, we report that peroxiredoxin (Prx) gene expression in castration-resistant prostate cancer and hydrogen peroxide-resistant cells was upregulated. Prx2 was overexpressed in castration-resistant prostate cancer at the mRNA and protein levels and was localized to the nucleus and cytoplasm. Overexpression of Prx2 increased AR transactivation, whereas Prx2 overexpression in the nucleus suppressed AR transactivation. These effects of Prx2 on AR activity were abolished by the introduction of function-disrupting mutations into Cys 51 and Cys 172. Silencing Prx2 reduced the expression of androgen-regulated genes and suppressed the growth of AR-expressing prostate cancer cells by inducing cell-cycle arrest at the G1 phase. Furthermore, Prx2 knockdown also suppressed cell growth in castration-resistant prostate cancer cells. These findings indicate that Prx2 is involved in the proliferation of AR-expressing prostate cancer cells by modulating AR activity. Designing therapeutics targeting Prx2 may offer a novel strategy for developing treatments for prostate cancer, including castration-resistant prostate cancer, which is dependent on AR signaling..
79. masaki shiota, Akira Yokomizo, Momoe Itsumi, Takeshi Uchiumi, Yasuhiro Tada, Yoohyun Song, Eiji Kashiwagi, Daisuke Masubuchi, Seiji Naito, Twist1 and Y-box-binding protein-1 promote malignant potential in bladder cancer cells, BJU international, 10.1111/j.1464-410X.2010.09810.x, 108, 2 B, 2011.07, What's known on the subject? and What does the study add? So far, transcription factor Twist1 has been reported to regulate Y-box binding protein-1 (YB-1) expression and play important roles in tumour growth, invasion and drug resistance. This study revealed that in bladder cancer cells, Twist1 regulated YB-1 transcription and both Twist1 and YB-1 promote malignant potentials including tumour growth, invasion and anticancer-drug resistance. Although the relation between Twist1/YB-1 signaling and cancer malignant potentials including tumour growth, invasion and anticancer-drug resistance have been suggested, their roles in bladder cancer remain unknown. This study revealed their functional importance in bladder cancer, indicating that both Twist1 and YB-1 are promising molecular targets in bladder cancer. Objective: To investigate the roles of Twist1 and Y-box binding protein-1 (YB-1) and their potential as therapeutic targets in bladder cancer (BC), as both have been suggested to play important roles in tumour growth, invasion and drug resistance. Materials and methods: Bladder cancer cell lines (TCCsup, UMUC3, T24 and KK47 cells) were used. Twist1 and YB-1 expression levels were assessed by luciferase reporter assay, quantitative real-time polymerase chain reaction (PCR) and western blot analysis. Tumour growth and cell cycle were analysed by cell proliferation assay and flow cytometry, respectively. Invasive and motile abilities were investigated by scratch-wound test and migration assay, respectively. Cytotoxicity assay was performed to determine drug sensitivity. Results: The findings showed that Twist1 regulated YB-1 expression in BC cells. Both Twist1 and YB-1 were involved in cell growth, invasion, motility and resistance to cisplatin and doxorubicin, but not to 5-fluorouracil (5-FU). Conclusion: The present study showed that Twist1 regulates YB-1 expression and that both Twist1 and YB-1 promote malignant potentials, including tumour growth, invasion and anti-cancer-drug resistance, indicating that both Twist1 and YB-1 are novel molecular targets in BC..
80. Shiota M, Yokomizo A, Kashiwagi E, Takeuchi A, Fujimoto N, Uchiumi T, Naito S. , Peroxiredoxin 2 in the nucleus and cytoplasm distinctly regulates androgen receptor activity in prostate cancer cells., Free Radic Biol Med, 51, 78-87, 2011.05.
81. Shiota M, Takeuchi A, Song Y, Yokomizo A, Kashiwagi E, Uchiumi T, Kuroiwa K, Tatsugami K, Fujimoto N, Oda Y, Naito S., Y-box binding protein-1 promotes castration-resistant prostate cancer growth via androgen receptor expression. , Endocr Relat Cancer., 18, 505-17, 2011.05.
82. Amamoto R, Yagi M, Song Y, Oda Y, Tsuneyoshi M, Naito S, Yokomizo A, Kuroiwa K, Tokunaga S, Kato S, Hiura H, Samori T, Kang D, Uchiumi T. , Mitochondrial p32/C1QBP is highly expressed in prostate cancer and is associated with shorter PSA relapse time after radical prostatectomy., Cancer Science, 2011.03.
83. Rie Amamoto, Mikako Yagi, Yoo Hyun Song, Yoshinao Oda, Masazumi Tsuneyoshi, Seiji Naito, Akira Yokomizo, Kentaro Kuroiwa, Shoji Tokunaga, Seiji Kato, Hisahide Hiura, Tomohiro Samori, Dongchon Kang, Takeshi Uchiumi, Mitochondrial p32/C1QBP is highly expressed in prostate cancer and is associated with shorter prostate-specific antigen relapse time after radical prostatectomy, Cancer Science, 10.1111/j.1349-7006.2010.01828.x, 102, 3, 639-647, 2011.03, Mitochondria are key organelles for ATP production and apoptosis. Therefore, impairment of mitochondria can modulate or accelerate cancer progression. p32, originally identified as a pre-mRNA splicing factor SF2/ASF-associated protein, is localized predominantly in the mitochondrial matrix and involved in mitochondria respiration. Recently, p32 was implicated in apoptosis and resultantly cancer progression. However, little is known about the expression and function of p32 in human tumors including prostate cancer. Here, we investigated the expression of p32 in 148 prostate carcinoma tissues by immunohistochemistry and found a positive correlation of p32 expression to clinicopathological parameters including follow-up data. p32 is highly expressed in prostate tumor samples and its expression is significantly associated with the Gleason score, pathological stage and relapse. For localized cancers, high p32 is a strong and independent predictor of clinical recurrence in multivariate analysis (P=0.01). In addition, p32 is overexpressed in the prostate cancer cell lines examined. The selective knockdown of p32 by RNA interference inhibits the growth of prostate cancer cell lines but not of a non-cancerous cell line. The p32 RNA interference decreases cyclin D1, increases p21 expression and causes a G1/S cell cycle arrest in prostate cancer cells. These data suggest that p32 is critical for prostate cancer cell proliferation and may be a novel marker of clinical progression in prostate cancer..
84. masaki shiota, Yoo Hyun Song, Akira Yokomizo, Keijiro Kiyoshima, Yasuhiro Tada, Hiroshi Uchino, Takeshi Uchiumi, Junichi Inokuchi, Yoshinao Oda, Kentaro Kuroiwa, Katsunori Tatsugami, Seiji Naito, Foxo3a suppression of urothelial cancer invasiveness through twist1, Y-box-binding protein 1, and E-cadherin regulation, Clinical Cancer Research, 10.1158/1078-0432.CCR-10-0376, 16, 23, 5654-5663, 2010.12, Purpose: Invasion and metastasis are key steps in the progression of urothelial cancer (UC) into a critical disease. Foxo3a is a member of the Foxo transcription factor family that modulates the expression of various genes. We aimed to elucidate the role of Foxo3a in UC invasion. Experimental Design: Foxo3a mRNA and protein expressions in UC samples were investigated by gene expression assays and immunohistochemistry, respectively. Foxo3a expression was compared with clinicopathologic characteristics and patient prognoses based on UC samples. Quantitative real-time polymerase chain reaction, Western blotting, and migration assays were also conducted in UC cells. Results: Foxo3a expression decreased in invasive UC; patients with low Foxo3a expression had poor disease-free survival, cancer-specific survival, and overall survival; Foxo3a knockdown in UC cells increased cellular motility. Foxo3a negatively regulated Twist1 and Y-box-binding protein 1 (YB-1), and positively regulated E-cadherin in KK47 and TCC sup cells that expressed Twist 1, but not in T24 cells that did not express Twist1. Foxo3a-associated acetyltransferase p300 and Foxo3a acetylation status also affected UC motility. Conclusion: The results of this study indicate that Foxo3a regulates motility of UC through negative regulation of Twist1 and YB-1, and through positive regulation of E-cadherin. This suggests that Foxo3a could act as an independent prognostic factor in UC and could represent a promising molecular target for cancer therapeutics..
85. Hiroto Izumi, Tetsuro Wakasugi, Shohei Shimajiri, Akihide Tanimoto, Yasuyuki Sasaguri, Eiji Kashiwagi, Yoshihiro Yasuniwa, Masaki Akiyama, Bin Han, Ying Wu, Takeshi Uchiumi, Tokuzo Arao, Kazuto Nishio, Ryuta Yamazaki, Kimitoshi Kohno, Role of ZNF143 in tumor growth through transcriptional regulation of DNA replication and cell-cycle-associated genes, Cancer Science, 10.1111/j.1349-7006.2010.01725.x, 101, 12, 2538-2545, 2010.12, The cell cycle is strictly regulated by numerous mechanisms to ensure cell division. The transcriptional regulation of cell-cycle-related genes is poorly understood, with the exception of the E2F family that governs the cell cycle. Here, we show that a transcription factor, zinc finger protein 143 (ZNF143), positively regulates many cell-cycle-associated genes and is highly expressed in multiple solid tumors. RNA-interference (RNAi)-mediated knockdown of ZNF143 showed that expression of 152 genes was downregulated in human prostate cancer PC3 cells. Among these ZNF143 targets, 41 genes (27%) were associated with cell cycle and DNA replication including cell division cycle 6 homolog (CDC6), polo-like kinase 1 (PLK1) and minichromosome maintenance complex component (MCM) DNA replication proteins. Furthermore, RNAi of ZNF143 induced apoptosis following G2/M cell cycle arrest. Cell growth of 10 lung cancer cell lines was significantly correlated with cellular expression of ZNF143. Our data suggest that ZNF143 might be a master regulator of the cell cycle. Our findings also indicate that ZNF143 is a member of the growing list of non-oncogenes that are promising cancer drug targets. (Cancer Sci 2010; 101: 2538-2545).
86. Izumi H, Wakasugi T, Shimajiri S, Tanimoto A, Sasaguri Y, Kashiwagi E, Yasuniwa Y, Akiyama M, Han B, Wu Y, Uchiumi T, Arao T, Nishio K, Yamazaki R, Kohno K, Role of ZNF143 in tumor growth through transcriptional regulation of DNA replication and cell-cycle-associated genes, Cancer Science, 101, 2538-2545, 2010.09.
87. Twist1 and Y-box-binding protein-1 promote malignant potential in bladder cancer cells., Twist1 and Y-box-binding protein-1 promote malignant potential in bladder cancer cells., BJU Int, 2010.09.
88. masaki shiota, Akira Yokomizo, Yasuhiro Tada, Takeshi Uchiumi, Junichi Inokuchi, Katsunori Tatsugami, Kentaro Kuroiwa, Ken Yamamoto, Narihito Seki, Seiji Naito, P300/CBP-associated factor regulates Y-box binding protein-1 expression and promotes cancer cell growth, cancer invasion and drug resistance, Cancer Science, 10.1111/j.1349-7006.2010.01598.x, 101, 8, 1797-1806, 2010.08, Twist1 has been proposed to have oncogenic properties. Although Twist1 was reported to interact with p300/CBP-associated factor (PCAF) and to inhibit the functions of PCAF, it remains unclear how PCAF affects the functions of Twist1, cell growth, invasive ability, and cellular sensitivity to anticancer agents. We found that PCAF, Twist1, and Y-box binding protein-1 (YB-1) expressions were elevated in cisplatin- and doxorubicin-resistant cancer cells. Luciferase reporter assays revealed that PCAF manipulation modulated YB-1 transcription in a Twist1-dependent manner. In addition, PCAF regulated the Twist1 intracellular localization and the Twist1 transcriptional activity through its acetylation function to the Twist1. Suppression of PCAF expression reduced YB-1 expression in human urothelial cancer KK47 cells. As a result, the cell growth and invasive ability of KK47 cells was retarded by PCAF knockdown, and PCAF knockdown rendered KK47 cells sensitive to cisplatin and doxorubicin, but not to 5-fluorouracil. The present data suggest that Twist1 and YB-1 as well as PCAF may be promising molecular therapeutic targets. (Cancer Sci 2010)..
89. masaki shiota, Masatoshi Eto, Akira Yokomizo, Yasuhiro Tada, ario takeuchi, Momoe Itsumi, Katsunori Tatsugami, Takeshi Uchiumi, Seiji Naito, Sensitivity of doxorubicin-resistant cells to sorafenib
Possible role for inhibition of eukaryotic initiation factor-2α phosphorylation, International Journal of Oncology, 10.3892/ijo-0000700, 37, 2, 509-517, 2010.08, Patients with advanced cancer including breast cancer, hepatocellular cancer and urothelial cancer frequently receive a chemotherapy regimen containing doxorubicin. However, doxorubicin-resistance is a major obstacle for cancer chemotherapy. Recently, several molecular-targeted agents have become available. Sorafenib (BAY 43-9006) is known to target multiple kinases and has demonstrated activity in renal cell and hepatocellular cancer. In this study, sorafenib was found to inhibit phosphorylation of the eukaryotic initiation factor-2α (eIF2α), induce cell cycle arrest at G2 phase and increase cellular apoptosis in doxorubicin-resistant human urothelial cell lines. An eIF2α kinase, PERK was responsible for eIF2α phosphorylation and PERK knockdown induced cellular apoptosis similar to sorafenib treatment in doxorubicin-resistant cancer cells. Furthermore, sorafenib sensitized doxorubicin-resistant cancer cells, but not their parental cells to oxidative stress exerted by both hydrogen peroxide and doxorubicin. In addition, PERK knockdown sensitized doxorubicin-resistant cancer cells to oxidative stress. In conclusion, PERK inhibition using sorafenib with or without doxorubicin might be a promising therapeutic approach for doxorubicin-resistant cancers retaining high phosphorylation levels of eIF2α..
90. Shiota M, Yokomizo A, Tada Y, Uchiumi T, Inokuchi J, Tatsugami K, Kuroiwa K, Yamamoto K, Seki N, Naito S., P300/CBP-associated factor regulates Y-box binding protein-1 expression and promotes cancer cell growth, cancer invasion and drug resistance., Cancer Science, 101, 1797-1806, 2010.07.
91. Shiota M, Song Y, Yokomizo A, Kiyoshima K, Tada Y, Uchino H, Uchiumi T, Inokuchi J, Oda Y, Kuroiwa K, Tatsugami K, Naito S, Foxo3a suppression of urothelial cancer invasiveness through Twist1, Y-box-binding protein 1, and E-cadherin regulation, Clin. Cancer Res, 16, 5654-5663, 2010.07.
92. Shiota M, Eto M, Yokomizo A, Tada Y, Takeuchi A, Itsumi M, Tatsugami K, Uchiumi T, Naito S, Sensitivity of doxorubicin-resistant cells to sorafenib: possible role for inhibition of eukaryotic initiation factor-2alpha phosphorylation, Int J Oncol, 37, 509-517, 2010.07.
93. Shiota, M. Eto, M. Yokomizo, A. Tada, Y. Takeuchi, A. Masubuchi, D. Inokuchi, J. Tatsugami, K. Kuroiwa, K. Uchiumi, T. Seki, N. Naito, S., Sorafenib with doxorubicin augments cytotoxicity to renal cell cancer through PERK inhibition, Int J Oncol, 36, 6, 1521-31, 2010.06.
94. Uchiumi, T. Ohgaki, K. Yagi, M. Aoki, Y. Sakai, A. Matsumoto, S. Kang, D., ERAL1 is associated with mitochondrial ribosome and elimination of ERAL1 leads to mitochondrial dysfunction and growth retardation, Nucleic Acids Res, 2010.05.
95. Shiota, M. Yokomizo, A. Tada, Y. Inokuchi, J. Tatsugami, K. Kuroiwa, K. Uchiumi, T. Fujimoto, N. Seki, N. Naito, S., Peroxisome proliferator-activated receptor gamma coactivator-1alpha interacts with the androgen receptor (AR) and promotes prostate cancer cell growth by activating the AR, Mol Endocrinol, 2010.05.
96. Shiota, M. Yokomizo, A. Kashiwagi, E. Tada, Y. Inokuchi, J. Tatsugami, K. Kuroiwa, K. Uchiumi, T. Seki, N. Naito, S., Foxo3a expression and acetylation regulate cancer cell growth and sensitivity to cisplatin, Cancer Sci, 2010.05.
97. Shiota, M. Song, Y. Yokomizo, A. Tada, Y. Kuroiwa, K. Eto, M. Oda, Y. Inokuchi, J. Uchiumi, T. Fujimoto, N. Seki, N. Naito, S., Human heterochromatin protein 1 isoform HP1{beta} enhances androgen receptor activity and is implicated in prostate cancer growth, Endocr Relat Cancer, 2010.05.
98. Shiota, M. Yokomizo, A. Masubuchi, D. Tada, Y. Inokuchi, J. Eto, M. Uchiumi, T. Fujimoto, N. Naito, S., Tip60 promotes prostate cancer cell proliferation by translocation of androgen receptor into the nucleus, Prostate, 70, 5, 540-54, 2010.04.
99. Shiota, M. Yokomizo, A. Tada, Y. Inokuchi, J. Kashiwagi, E. Masubuchi, D. Eto, M. Uchiumi, T. Naito, S., Castration resistance of prostate cancer cells caused by castration-induced oxidative stress through Twist1 and androgen receptor overexpression, Oncogene, 29, 2, 237-50, 2010.01.
100. Fukuoh, A. Ohgaki, K. Hatae, H. Kuraoka, I. Aoki, Y. Uchiumi, T. Jacobs, H. T.Kang, D., DNA conformation-dependent activities of human mitochondrial RNA polymerase, Genes Cells, 14, 8, 1029-42, 2009.08.
101. Niina I, Uchiumi T, Izumi H,Torigoe T, Wakasugi T, Igarashi T, Miyamoto N, Onitsuka T, Shiota M, Okayasu R, Chijiiwa K, and Kohno K, DNA topoisomerase inhibitor, etoposide, enhances GC-box-dependent promoter activity via Sp1 phosphorylation, Cancer Science, 2007.10.
102. Momii Y, Izumi H, Shiota M, Onistuka T, Abe T, Kobayasi H, Miyamoto N, Uchiumi T, and Kohno K, p73gamma transactivates the p21 promoter through preferential interaction with the p300/CBP-associated factor in human prostate cancer cells., Oncology Report, 18, 411-416, 2007.10.
103. Uramoto H, Uchiumi T, Izumi H, Kohno K, Oyama T, Sugio K, and Yasumoto K, A new mechanism for primary resistance to gefitinib in lung adenocarcinoma: the role of a novel G796A mutation in exon 20 of EGFR, Anticancer Res,, 27, 2297-2303, 2007.10.
104. Wakasugi,T., Izumi,H., Uchiumi,T., Suzuki,H., Arao,T., Nishio,K. and Kohno,K, ZNF143 interacts with p73 and is involved in cisplatin resistance through the transcriptional regulation of DNA repair genes., Oncogene, 26: 5194-5203, 2007.09.
105. Igarashi,T., Izumi,H., Uchiumi,T., Nishio,K., Arao,T., Uramoto,H., Sugio,K., Yasumoto,K., Sasaguri,Y., Wang,KY., Otsuji,Y. and Kohno,K., Clock/ATF4 transcription system regulates multidrug resistance in human cancer cell line., Oncogene, 26, 4749-4760, 2007.06.
106. Uchiumi,T., Fotovati,A., Sasaguri,T., Shibahara,K., Shimada,T., Fukuda,T., Nakamura,T., Izumi,H., Tsuzuki,T., Kuwano,M. and Kohno,K, YB-1 is important for an early stage embryonic development: neural tube formation and cell proliferation. , J. Biol.Chem, 281: 40440-40449, 2006.11.
107. Yamashita,F., Ohtani,H., Koyabu,N., Ushigome,F., Satoh,H., Murakami,H., Uchiumi,T., Nakamura,T., Kuwano,M., Tsujimoto,M. and Sawada,Y., Inhibitory effects of angiotensin II receptor antagonists and leukotriene receptor antagonists on the transport of human organic anion transporter 4. , J. Pharm. Pharmacol., 2006.06.
108. Kohno,K., Matsuki,Y., Tanimoto,A., Izumi,H., Uchiumi,T., Kohno,K., Shimajiri,S. and Sasaguri,Y., Expression of Y-box-binding protein dbpC/contrin, a potentially new cancer/testis antigen., Br. J. Cancer, 94: 710-716, 2006.05.
109. Yoshida,T., Izumi,H., Uchiumi,T., Sasaguri,Y., Tanimoto,A., Matsumoto,T., Naito,S. and Kohno,K., Expression and cellular localization of dbpC/Contrin in germ cell tumor cell lines., Biochim. Biophys. Acta, 1759: 80-88, 2006.03.
110. Oda,Y., Saito,T., Tateishi,N., Ohishi,Y., Tamiya,S.,Yamamoto,H.,Yokoyama,R., Uchiumi,T., Iwamoto,Y., Kuwano,M. and Tsuneyoshi,M, ATP-binding cassette superfamily transporter gene expression in human soft tissue sarcoma., Int. J. Cancer, 10.1002/ijc.20589, 114, 6, 854-862, 2005.11.
111. Sata,R., Ohtani,H., Tsujimoto,M., Murakami,H., Koyabu,N., Nakamura,T., Uchiumi,T., Kuwano,M., Nagata,H., Tsukimori,K., Nakano,H. and Sawada,Y, Functional analysis of organic cation transporter 3 expressed in human placenta. , J. Pharmacol. Exp. Ther., 10.1124/jpet.105.086827, 315, 2, 888-895, 2005.10.
112. Pulaski,L., Kania,K., Ratajewski,M., Uchiumi,T., Kuwano,M. and Bartosz,G. , Differential regulation of the human MRP2 and MRP3 gene expression by glucocorticoids. , J. Steroid Biochem. Mol. Biol., 10.1016/j.jsbmb.2005.03.004, 96, 3-4, 229-234, 96: 229-234, 2005.08.
113. Yasen,M., Kajino,K., Kano,S., Tobita,H., Yamamoto,J., Uchiumi,T., Kon,S., Maeda,M., Obulhasim,G., Arii,S. and Hino,O., The up-regulation of Y-box binding proteins (DNA binding protein A and Y-box binding protein-1) as prognostic markers of hepatocellular carcinoma., Clin. Cancer Res, 11: 7354-7361, 2005.07.
114. Pulaski,L., Szemraj,J., Uchiumi,T., Kuwano,M. and Bartosz,G., Transcriptional upregulation of the human MRP2 gene expression by serine/threonine protein kinase inhibitors., J. Biol. Regul. Homeost. Agents, 19, 3-4, 113-119, 19: 113-119, 2005.05.
115. Honda,Y., Ushigome,F., Koyabu,N., Morimoto,S., Shoyama,Y., Uchiumi,T., Kuwano,M., Ohtani H. and Sawada Y., Effects of grapefruit juice and orange juice components on P-glycoprotein- and MRP2-mediated drug efflux., Br. J. Pharmacol., 10.1038/sj.bjp.0706008, 143, 7, 856-864, 143: 856-864, 2004.11.
116. Shibahara,K., Uchiumi,T., Fukuda,T., Kura,S., Tominaga,Y., Maehara,Y., Kohno,K., Nakabeppu,Y., Tsuzuki,T. and Kuwano,M., Targeted disruption of one allele of the Y-box binding protein-1 (YB-1) gene in mouse embryonic stem cells and increased sensitivity to cisplatin and mitomycin C, Cancer Sci., 10.1111/j.1349-7006.2004.tb03214.x, 95, 4, 348-353, 95: 348-353, 2004.07.
117. Hisaeda,K., Inokuchi,A., Nakamura,T., Iwamoto,Y., Kohno,K., Kuwano,M. and Uchiumi,T. , Interleukin-1ß represses MRP2 gene expression through inactivation of interferon regulatory factor 3 in HepG2 cells., Hepatology, 10.1002/hep.20216, 39, 6, 1574-1582, 39: 1574-1582, 2004.04.
118. Fukuda, T., Ashizuka, M., Nakamura, T., Shibahara,K., Maeda, K., Izumi, I., Kohno, K., Kuwano, M.,and Uchiumi,T.,, Characterization of 5'untranslated region of YB-1 mRNA and plausible regulation of translation., Nucleic Acid Res, 2004.01.
119. Ushigome, F., Koyabu, N., Satoh, S., Tsukimori, K., Nakano, H., Nakamura, T., Uchiumi, T., Kuwano, M., Ohtani, H., and Sawada, Y, Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles., Pharm Res, 10.1023/A:1022290523347, 20, 1, 38-44, 20: 38-44, 2003.01.
120. Kuwano, M., Uchiumi, T., Hayakawa, H., Ono, M., Wada, M., Izumi, H., and Kohno, K, The basic and clinical implications of ABC transporters, Y-box-binding protein-1 (YB-1) and angiogenesis-related factors in human malignancies., Cancer Sci, 10.1111/j.1349-7006.2003.tb01344.x, 94, 1, 9-14, 94: 9-14, 2003.01.
121. Konno, T., Ebihara, T., Hisaeda, K., Uchiumi, T., Nakamura, T., Shirakusa, T., Kuwano, M., and Wada, M., Identification of domains participating in the substrate specificity and subcellular localization of the multidrug resistance proteins MRP1 and MRP2., J Biol Chem, 10.1074/jbc.M302868200, 278, 25, 22908-22917, 278: 22908-22917, 2003.01.
122. Nagashige M, Ushigome F, Koyabu N, Hirata K, Kawabuchi M, Hirakawa T, Satoh, Basal membrane localization of MRP1 in human placental trophoblast., Placenta, 10.1016/S0143-4004(03)00170-X, 24, 10, 951-958, 24: 951-958, 2003.01.
123. Kohno K, Izumi H, Uchiumi T, Ashizuka M, and Kuwano M, The pleiotropic functions of the Y-box-binding protein, YB-1, Bioessays, 10.1002/bies.10300, 25, 7, 691-698, 25: 691-698, 2003.01.
124. Zhu, H. Chang,B.-D. Uchiumi, T. and Roninson, I. B, Identification of promoter elements responsible for transcriptional inhibition of PLK1 and Topoisomerase IIa genes by p21WAF1/CIP1/SDI1., Cell cycle,, 1: 59-66, 2002.01.
125. Yamamoto, C., Murakami, H., Koyabu, N., Takanaga, H., Matsuo, H., Uchiumi, T., Kuwano, M., Naito, M., Tsuruo, T., Ohtani, H., and Sawada, Y, Contribution of P-glycoprotein to efflux of ramosetron, a 5-HT3 receptor antagonist, across the blood-brain barrier., J Pharm Pharmacol, 10.1211/002235702320266208, 54, 8, 1055-1063, 54: 1055-1063., 2002.01.
126. Uramoto, H., Izumi, H., Ise, T., Tada, M., Uchiumi, T., Kuwano, M., Yasumoto, K., Funa, K., and Kohno, K., p73 Interacts with c-Myc to regulate Y-box-binding protein-1 expression, J Biol Chem,, 10.1074/jbc.M200266200, 277, 35, 31694-31702, 277: 31694-31702, 2002.01.
127. Kakehi, M., Koyabu, N., Nakamura, T., Uchiumi, T., Kuwano, M., Ohtani, H., and Sawada, Y, Functional characterization of mouse cation transporter mOCT2 compared with mOCT1., Biochem Biophys Res Commun, 10.1016/S0006-291X(02)00926-9, 296, 3, 644-650, 296: 644-650, 2002.01.
128. Hayakawa, H., Uchiumi, T., Fukuda, T., Ashizuka, M., Kohno, K., Kuwano, M., and Sekiguchi, M, Binding Capacity of Human YB-1 Protein for RNA Containing 8-Oxoguanine., Biochemistry, 10.1021/bi0201872, 41, 42, 12739-12744, 41: 12739-12744, 2002.01.
129. Ashizuka, M., Fukuda, T., Nakamura, T., Shirasuna, K., Iwai, K., Izumi, H., Kohno, K., Kuwano, M., and Uchiumi, T., Novel translational control through an iron-responsive element by interaction of multifunctional protein YB-1 and IRP2., Mol Cell Biol, 10.1128/MCB.22.18.6375-6383.2002, 22, 18, 6375-6383, 22: 6375-6383, 2002.01.
130. Hashimoto K.,Uchiumi T., Konno T., Ebihara T., Nakamura T., Wada M., Sakisaka S., Maniwa F., Amachi T., Ueda K., and Kuwano M., Trafficking and functional defects by mutations of the ATP-binding domains in MRP2 in patients with Dubin-Johnson syndrome., Hepatology, 10.1053/jhep.2002.36368, 36, 5, 1236-1245, 36:1236-1245, 2002.01.
131. Ohishi Y., Oda Y., Uchiumi T., Kobayashi H., Hirakawa T., Miyamoto S.,Kinukawa N.,Nakano H., Kuwano M., and Tsuneyoshi M., ATP-binding cassette superfamily transporter gene expression in human primary ovarian carcinoma., Clin Cancer Res, 8, 12, 3767-3775, 8:3767-3775, 2002.01.
132. Haga, S., Hinoshita, E., Ikezaki, K., Fukui, M., Scheffer, G. L., Uchiumi, T. and Kuwano, M., Involvement of the multidrug resistance protein 3 in drug sensitivity and its expression in human glioma., Jpn. J. Cancer Res, 92, 2, 211-219, 92: 211-219, 2001.01.
133. Izumi, H., Imamura, T., Nagatani, G., Ise, T., Murakami, T., Uramoto, H., Torigoe, T., Ishiguchi, H., Nomoto, M., Okamoto, T., Uchiumi, T., Kuwano, M., Funa, K. and Kohno, K., YB-1 binds preferentially to single-stranded nucleic acid and exhibit 3'-5' exonuclease activity., Nucl. Acids Res., 29: 1200-1207, 2001.01.
134. Hinoshita, E., Taguchi, K., Inokuchi, A., Uchiumi, T., Kinukawa, N., Shimada, M., Tsuneyoshi, M., Sugimachi, K. and Kuwano, M., Decreased expression of an ATP-binding cassette transporter, MRP2, in human livers with hepatitis C virus infection., J. Hepatology, 10.1016/S0168-8278(01)00216-1, 35, 6, 765-773, 35,765-773, 2001.01.
135. Kawakami, A., Tomofumi, M., Higuchi, R., Uchiumi, T., Kuwano, M. and Van Soest, R. W. N., Structure of a novel multidrug resistance modulator, irciniasulfonic acid, isolated from a marin sponge, Ircinia sp., Tetrahedron Lett., 10.1016/S0040-4039(01)00426-9, 42, 19, 3335-3337, 42: 3335-3337, 2001.01.
136. Shibahara, K., Sugio, K., Osaki, T., Uchiumi, T., Maehara, Y., Kohno, K., Yasumoto, K., Sugimati, K. and Kuwano, M., Nuclear expression of the Y-box binding protein, YB-1, as a novel marker of disease progression in non-small-cell lung cancer., Clinical Cancer Res., 7, 10, 3151-3155, 7: 3151-3155, 2001.01.
137. Inokuchi, A., Hinoshita, E., Wada, M., Iwamoto, Y., Kohno, K., Kuwano, M. and Uchiumi, T., Enhanced expression of Human Multidrug resistance protein 3 by bile salt in human enterocytes: a transcriptional control of plausible bile acid transporter., J. Biol. Chem., 10.1074/jbc.M104612200, 276, 50, 46822-46829, 276: 46822-46829, 2001.01.
138. Okamoto, M., Ono, M., Uchiumi, T., Ueno, H., Kohno, K., Sugimachi, K. and Kuwano, M., Up-regulation of thrombospondin-1 gene by epidermal growth factor and transforming growth facter b in human cancer cells-transcriptional activation and messenger RNA stabilization., Biochim. Biophys. Acta, 93595, 2001.01.
139. Hinoshita, E., Uchiumi, T., Taguchi, K., Kinukawa, N., Tsuneyoshi, M., Maehara, Y., Sugimachi, K. and Kuwano, M., Increased expression of an ATP-binding cassette superfamily transporter, multidrug resistance protein 2, in human colorectal carcinomas., Clin Cancer Res, 6, 6, 2401-2407, 6:2401-2407, 2000.01.
140. Okamoto, T., Izumi, H., Imamura, T., Takano, H., Ise, T, Uchiumi, T., Kuwano, M. and Kohno, K., Direct interaction of p53 with the Y-box binding protein, YB-1: a mechanism for regulation of human gene expression., Oncogene, 10.1038/sj.onc.1204029, 19, 54, 6194-6202, 19: 6194-6202, 2000.01.
141. Ushigome, F., Takanaga, H., Matsuo, H., Yanai, S., Tsukimori, K., Nakano, H., Uchiumi, T., Nakamura, T., Kuwano, M., Ohtani, H., and Sawada, Y., Human placental transport of vinblastine, vincristine, digoxin and progesterone: contribution of P-glycoprotein., Eur. J. Pharmacol, 10.1016/S0014-2999(00)00743-3, 408, 1, 1-10, 408: 1-10, 2000.01.
142. Ohga,T., Uchiumi,T., Makino, Y., Koike,K., Wada,M., Kuwano, M. and Kohno,K., Direct involvement of the Y-box binding protein YB-1 in genotoxic stress-induced activation of the human multidrug resistance 1 gene., J. Biol. Chem., 10.1074/jbc.273.11.5997, 273, 11, 5997-6000, 273: 5997-6000, 1999.01.
143. Toh, S., Wada, M., Uchiumi, T., Inokuchi, A., Makino, Y., Horie, H., Adachi, Y., Sakisaka, S. and Kuwano, M., Genomic structure of the canalicular multispecitic organic anion transporter (cMOAT) gene and mutations in the ATP binding cassette region in Dubin-Johnson syndrome., Am. J. Human Genet., 10.1086/302292, 64, 3, 739-746, 64: 739-746, 1999.01.
144. Ise, T., Nagatani, G., Imamura, T., Kato, K., Takano, H., Nomoto, M., Izumi, H., Ohmori, H., Okamoto, T., Ohga, T., Uchiumi, T., Kuwano, M. and Kohno, K., Transcription factor Y box-binding protein-1 binds preferentially to cisplatin-modified DNA and interacts with proliferating cell nuclear antigen., Cancer Res, 59, 2, 342-346, 59: 342-346, 1999.01.
145. Chen, Z., Kawabe, T., Ono, M., Aoki, S., Sumizawa, T., Furukawa, T., Uchiumi, T., Wada, M., Kuwano, M. and Akiyama, S., Effect of multidrug resistance-reversing agents on transporting activity of human canalicular multispecific organic anion transporter., Molec. Pharmacology, 56, 6, 1219-1228, 56: 1219-1228, 1999.01.
146. Kawabe, T., Chen, Z., Wada, M., Uchiumi, T., Ono, M., Akiyama, S. and Kuwano, M., Enhanced transport of anticancer agents and leukotriene C4 by the human canalicular multispecific organic anion transporter (cMOAT/MRP2)., FEBS Lett., 10.1016/S0014-5793(99)00979-5, 456, 2, 327-331, 456: 327-331, 1999.01.
147. Tanaka,T., Uchiumi,T., Hinoshita,E., Inokuchi,A., Toh,S., Wada,M., Takano,H., Kohno,K., and Kuwano,M., The human multispecific resistance protein2 gene : functional characterization of 5'-flanking region and expression in hepatic cells., Hepatology, 30: 1507-1512, 1999.01.
148. Sibao, K., Takano, H., Nakamura, Y., Okazaku, K., Nagata, N., Izumi, H., Uchiumi, T., Kuwano, M., Kohno, K. and Itoh, H., Enhanced coexpression of YB-1 and DNA topoisomerase IIαgenes in human colorectal carcinomas., Int. J.Cancer, 10.1002/(SICI)1097-0215(19991210)83:6<732::AID-IJC6>3.3.CO;2-R, 83, 6, 732-737, 83: 732-73, 1999.01.
149. Toh,S., Nakamura,T., Ohga, T., Koike,K., Uchiumi,T., Wada,M., Kuwano,M. and Kohno,K., Genomic organization of the human Y-box protein (YB-1) gene., Gene, 10.1016/S0378-1119(97)00570-2, 206, 1, 93-97, 206: 93-97, 1998.01.
150. Wada,M., Toh, S., Taniguchi, K., Uchiumi,T. , Kohno,K., Yoshida, I., Kimura,A., Sakisaka, S., Adachi, Y. and Kuwano,M., Mutation in the canalicular multispecific organic anion transporter (cMOAT) gene, a novel ABC transporter, in patients with hyperbilirubinemia II/Dubin-Johnson syndrome., Human Molec. Genet., 10.1093/hmg/7.2.203, 7, 2, 203-207, 7: 203-207, 1998.01.
151. Torigoe, K., Harada, T., Kusaba, H., Uchiumi, T., Kohno, K., E.D.Green, S. W.Scherer, L-C.Tsui, D.Schlessinger, Kuwano,M. and Wada, M., Localization of 67 exons on YAC contigs spanning 1.5 megabases around the multidrug resistance gene region of human chromosome 7q21.1., Genomics, 10.1006/geno.1997.5200, 49, 1, 14-22, 48:14-22, 1998.01.
152. Furukawa,M., Uchiumi,T., Nomoto,M., Takano,H., Morimoto,R-I., Naitoh,S., Kuwano,M. and Kohno,K., The role of an inverted CCAAT element in transcriptional activation of the human DNA topoisomeraseIIα gene in response to heat shock., J. Biol. Chem., 10.1074/jbc.273.17.10550, 273, 17, 10550-10555, 273: 10550-10555, 1998.01.
153. Oda,Y., Sakamoto,A., Shinohara,N., Ohga,T., Uchiumi,T., Kohno,K., Tsuneyoshi,M., Kuwano,M. and Iwamoto,Y., Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human osteosarcoma., Clin Cancer Res, 4, 9, 2273-2277, 4:2273-2277, 1998.01.
154. Uchiumi, T., Hinoshita, E., Haga, S., Nakamura, T., Tanaka, T., Toh, S., Furukawa, M., Kawabe, T., Wada, M., Kagotani, K., Okumura, K., Kohno, K., Akiyama, S. and Kuwano, M., Isolation of a novel human canalicular multispecific organic anion transporter, cMOAT2/MRP3, and its expression in cisplatin-resistant cancer cells with decreased ATP-dependent drug transport., Biochem. Biophys. Res. Commun, 10.1006/bbrc.1998.9546, 252, 1, 103-110, 252: 103-110, 1998.01.
155. Uchiumi, T., Longo, D. and Ferris, D, Cell cycle regulation of PLK promoter., J. Bio. Chem, 272,9166-9174, 1997.01.
156. Rie Amamoto, Takeshi Uchiumi, Mikako Yagi, Keisuke Monji, Song, Yoshinao Oda, Shiota M, Akira Yokomizo, Seiji Naito, Dongchon Kang, The Expression of Ubiquitous Mitochondrial Creatine Kinase Is Downregulated as Prostate Cancer Progression, JOURNAL OF CANCER, 10.7150/jca.13207, 7, 1, 50-59.