Kyushu University Academic Staff Educational and Research Activities Database
List of Papers
Tamotsu Kiyoshima Last modified date:2021.09.19

Professor / Maxillofacial Diagnostic & Surgical Sciences / Department of Dental Science / Faculty of Dental Science


Papers
1. Akane Inoue, Tamotsu Kiyoshima, Keigo Yoshizaki, Chihiro Nakatomi, Mitsushiro Nakatomi, Hayato Ohshima, Masashi Shin, Jing Gao, Kanji Tsuru, Koji Okabe, Ichiro Nakamura, Hiroaki Honda, Miho Matsuda, Ichiro Takahashi, Eijiro Jimi, Deletion of epithelial cell-specific p130Cas impairs the maturation stage of amelogenesis, Bone, in press, 2021.09, Amelogenesis consists of secretory, transition, maturation, and post-maturation stages, and the morphological changes of ameloblasts at each stage are closely related to their function. p130 Crk-associated substrate (Cas) is a scaffold protein that modulates essential cellular processes, including cell adhesion, cytoskeletal changes, and polarization. The expression of p130Cas was observed from the secretory stage to the maturation stage in ameloblasts. Epithelial cell-specific p130Cas-deficient (p130Casepi-) mice exhibited enamel hypomineralization with chalk-like white mandibular incisors in young mice and attrition in aged mouse molars. A micro-computed tomography analysis and Vickers micro-hardness testing showed thinner enamel, lower enamel mineral density and hardness in p130Casepi- mice in comparison to p130Casflox/flox mice. Scanning electron microscopy, and an energy dispersive X-ray spectroscopy analysis indicated the disturbance of the enamel rod structure and lower Ca and P contents in p130Casepi- mice, respectively. The disorganized arrangement of ameloblasts, especially in the maturation stage, was observed in p130Casepi- mice. Furthermore, expression levels of enamel matrix proteins, such as amelogenin and ameloblastin in the secretory stage, and functional markers, such as alkaline phosphatase and iron accumulation, and Na+/Ca2++K+-exchanger in the maturation stage were reduced in p130Casepi- mice. These findings suggest that p130Cas plays important roles in amelogenesis..
2. Jing Gao, Ryusuke Muroya, Fei Huang, Kengo Nagata, Masashi Shin, Ryoko Nagano, Yudai Tajiri, Shinsuke Fujii, Takayoshi Yamaza, Kazuhiro Aoki, Yukihiko Tamura, Mayuko Inoue, Sakura Chishaki, Toshio Kukita, Koji Okabe, Miho Matsuda, Yoshihide Mori, Tamotsu Kiyoshima, Eijiro Jimi, Bone morphogenetic protein induces bone invasion of melanoma by epithelial–mesenchymal transition via the Smad1/5 signaling pathway, Laboratory Investigation, 2021.09, Oral malignant melanoma, which frequently invades the hard palate or maxillary bone, is extremely rare and has a poor prognosis. Bone morphogenetic protein (BMP) is abundantly expressed in bone matrix and is highly expressed in malignant melanoma, inducing an aggressive phenotype. We examined the role of BMP signaling in the acquisition of an aggressive phenotype in melanoma cells in vitro and in vivo. In five cases, immunohistochemistry indicated the phosphorylation of Smad1/5 (p-Smad1/5) in the nuclei of melanoma cells. In the B16 mouse and A2058 human melanoma cell lines, BMP2, BMP4, or BMP7 induces morphological changes accompanied by the downregulation of E-cadherin, and the upregulation of N-cadherin and Snail, markers of epithelial–mesenchymal transition (EMT). BMP2 also stimulates cell invasion by increasing matrix metalloproteinase activity in B16 cells. These effects were canceled by the addition of LDN193189, a specific inhibitor of Smad1/5 signaling. In vivo, the injection of B16 cells expressing constitutively activated ALK3 enhanced zygoma destruction in comparison to empty B16 cells by increasing osteoclast numbers. These results suggest that the activation of BMP signaling induces EMT, thus driving the acquisition of an aggressive phenotype in malignant melanoma..
3. Sugiyama G, Ohyama Y, Yamada T, Ishii K, Kumamaru W, Sumimoto Y, Kiyoshima T, Niiro H, Mori Y., Other iatrogenic immunodeficiency-associated lymphoproliferative disorders of the oral floor induced by methotrexate and tofacitinib: A case report, JOURNAL OF ORAL AND MAXILLOFACIAL SURGERY MEDICINE AND PATHOLOGY, 10.1016/j.ajoms.2020.12.002, 33, 3, 297-301, 2021.05, Other iatrogenic immunodeficiency-associated lymphoproliferative disorders (0IIA-LPDs), which are uncontrolled lymphoid proliferation or lymphoma induced by immunosuppressive medication, arise mainly in patients with rheumatoid arthritis (RA). The vast majority of OIIA-LPDs are caused by methotrexate (MTX), and the reported incidence rates in the oral and maxillofacial regions are similar to those in other organs. In addition, tofacitinib (TFC), which is a Janus kinase inhibitor, has been clinically applied for the immunosuppressive treatment of RA in recent years. Herein, we present a case of OIIA-LPD of the oral floor in a patient with RA who had been treated with TFC and MTX. Clinical and pathological findings suggested that inflammatory stomatitis and/or infective response by oral bacteria promoted lymphoid activation in the oral mucosa, which could potentially become malignant. This case report indicates that OIIA-LPD might result from the combined usage of MTX and TFC, and a synergistic influence between MTX and TFC on immunosuppression may lead to diversification of OIIA-LPDs. The findings suggest the difficulty in controlling complications of RA and the importance of understanding the mechanisms of immunosuppressive treatments..
4. T Hattori, S Kawano, S Tanaka, R Matsubara, T Sakamoto, Y Hashiguchi, N Kaneko, Y Mikami, M Morioka, Y Maruse, R Kitamura, E Hamada, M Hiwatashi, K Oobu, T Kiyoshima, S Nakamura, Elevated Expression of Protease-Activated Receptor 1 via ΔNp63 Down-Regulation Contributes to Nodal Metastasis in Oral Squamous Cell Carcinoma, Int J Oral Maxillofac Surg, 10.1016/j.ijom.2020.04.021., 50, 2, 163-170, 2021.02, Protease-activated receptor 1 (PAR1) is known as a thrombin receptor. Recent studies have reported PAR1 expression in various malignancies; however, its role in oral squamous cell carcinoma (OSCC) requires clarification. A previous study showed that down-regulation of ΔNp63, a homolog of p53, augments PAR1 expression in OSCC. In the present study, the association of PAR1 expression with clinicopathological findings in OSCC was examined retrospectively. Expression of PAR1, thrombin, and ΔNp63 was examined immunohistochemically in OSCC specimens. Patients were divided into three groups based on the expression pattern of PAR1 at the invasive front: group A, PAR1-negative in both cancer and stromal cells; group B, positive in stromal cells but negative in cancer cells; group C, positive in both cancer and stromal cells. Histologically high-grade tumours were significantly more common in group C. Patients in group C had the highest incidence rate of nodal metastasis (P<0.001) and a lower survival rate (P=0.085) than those in the other groups. At the invasive front, in group C, thrombin was expressed but ΔNp63 expression was weak. These results indicate that increased PAR1 expression in both cancer and stromal cells could be a useful predictive marker of nodal metastasis and that ΔNp63 is involved in regulating PAR1 expression..
5. Kana Hasegawa, Shinsuke Fujii, Shinji Matsumoto, Yudai Tajiri, Akira Kikuchi, Tamotsu Kiyoshima, YAP signaling induces PIEZO1 to promote oral squamous cell carcinoma cell proliferation, J Pathol, 10.1002/path.5553, 253, 1, 80-93, 2021.01, Most cancer cells are exposed to altered extracellular environments, such as an increase in extracellular matrix (ECM) stiffness and soluble signals consisting of growth factors and cytokines. It is therefore conceivable that changes in tumor extracellular environments affect tumor cell behavior. The Hippo pathway reportedly responds to the extracellular environment and regulates the nuclear localization of the transcription co-activator, yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ). Inactivation of the Hippo pathway with nuclear translocation of YAP/TAZ stimulates cell proliferation. Its pathway also regulates gene expression, but the precise molecule(s) meditating the cell-proliferating effect of YAP signaling on oral squamous cell carcinoma (OSCC) is unclear. First, we examined the effects of YAP signaling on OSCC tumorigenesis. Loss-of-function experiments using siRNA or an inhibitor, and immunohistochemical analyses of tissue specimens obtained from OSCC patients demonstrated that YAP signaling was involved in OSCC cell proliferation. Second, we identified Piezo-type mechanosensitive ion channel component 1 (PIEZO1), a Ca2+ channel, as a transcriptional target of YAP signaling and showed that elevated PIEZO1 was required for PIEZO1 agonist-dependent Ca2+ entry and cell proliferation in OSCC cells. Experiments using three-dimensional and suspension culture revealed that PIEZO1 was involved in OSCC cellular growth. Finally, YAP overexpression in the nucleus and/or cytoplasm was immunohistochemically detected in tumor lesions with frequent expression of both PIEZO1 and Ki-67, but not in non-tumor regions of OSCC specimens. These results suggest that the YAP/PIEZO1 axis promotes OSCC cell growth.
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6. Tatsuki Yaginuma, Jing Gao, Kengo Nagata, Ryusuke Muroya, Huang Fei, Haruki Nagano, Sakura Chishaki, Takuma Matsubara, Shoichiro Kokabu, Kou Matsuo, Tamotsu Kiyoshima, Izumi Yoshioka, Eijiro Jimi , p130Cas Induces Bone Invasion by Oral Squamous Cell Carcinoma by Regulating Tumor Epithelial-Mesenchymal Transition and Cell Proliferation, Carcinogenesis, 10.1093/carcin/bgaa007., 2020.07, Bone invasion is a critical factor in determining the prognosis of oral squamous cell carcinoma (OSCC) patients. Transforming growth factor β (TGF-β) is abundantly expressed in the bone matrix and is involved in the acquisition of aggressiveness by tumors. TGF-β) is also important to cytoskeletal changes during tumor progression. In this study, we examined the relationship between TGF-β signaling and cytoskeletal changes during bone invasion by OSCC. Immunohistochemical staining of OSCC samples from 5 patients showed the expression of p130Cas (Crk-associated substrate) in the cytoplasm and phosphorylated Smad3 expression in the nucleus in OSCC cells. TGF-β1 induced the phosphorylation of Smad3 and p130Cas as well as epithelial-mesenchymal transition (EMT) accompanied by the downregulation of the expression of E-cadherin, a marker of epithelial cells, and the upregulation of the expression of N-cadherin, or Snail, a marker of mesenchymal cells, in human HSC-2 cells and mouse SCCVII cells. SB431542, a specific inhibitor of Smad2/3 signaling, abrogated the TGF-β1-induced phosphorylation of p130Cas and morphological changes. Silencing p130Cas using an shRNA or siRNA in SCCVII cells suppressed TGF-β1-induced cell migration, invasion, EMT, and matrix metalloproteinase-9 (MMP-9) production. Compared with control SCCVII cells, SCCVII cells with silenced p130Cas strongly suppressed zygomatic and mandibular destruction in vivo by reducing the number of osteoclasts, cell proliferation and MMP-9 production. Taken together, these results showed the expression of TGF-β/p130Cas might be a new target for the treatment of OSCC bone invasion..
7. Daigaku Hasegawa, Kana Hasegawa, Hiroshi Kaneko, Shinichiro Yoshida, Hiromi Mitarai, Mai Arima, Atsushi Tomokiyo, Sayuri Hamano, Hideki Sugii, Naohisa Wada, Tamotsu Kiyoshima, Hidefumi Maeda, MEST Regulates the Stemness of Human Periodontal Ligament Stem Cells, Stem Cells Int, 10.1155/2020/9672673, 2020, 2020.07, Periodontal ligament (PDL) stem cells (PDLSCs) have been reported as a useful cell source for periodontal tissue regeneration. However, one of the issues is the difficulty of obtaining a sufficient number of PDLSCs for clinical application because very few PDLSCs can be isolated from PDL tissue of donors. Therefore, we aimed to identify a specific factor that converts human PDL cells into stem-like cells. In this study, microarray analysis comparing the gene profiles of human PDLSC lines (2-14 and 2-23) with those of a cell line with a low differentiation potential (2-52) identified the imprinted gene mesoderm-specific transcript (MEST). MEST was expressed in the cytoplasm of 2-23 cells. Knockdown of MEST by siRNA in 2-23 cells inhibited the expression of stem cell markers, such as CD105, CD146, p75NTR, N-cadherin, and NANOG; the proliferative potential; and multidifferentiation capacity for osteoblasts, adipocytes, and chondrocytes. On the other hand, overexpression of MEST in 2-52 cells enhanced the expression of stem cell markers and PDL-related markers and the multidifferentiation capacity. In addition, MEST-overexpressing 2-52 cells exhibited a change in morphology from a spindle shape to a stem cell-like round shape that was similar to 2-14 and 2-23 cell morphologies. These results suggest that MEST plays a critical role in the maintenance of stemness in PDLSCs and converts PDL cells into PDLSC-like cells. Therefore, this study indicates that MEST may be a therapeutic factor for periodontal tissue regeneration by inducing PDLSCs..
8. Mizuki Sakamoto, Masafumi Moriyama, Mayumi Shimizu, Akira Chinju, Keita Mochizuki, Ryusuke Munemura, Keiko Ohyama , Takashi Maehara, Kenichi Ogata, Miho Ohta, Masaki Yamauchi, Noriko Ishiguro, Mayu Matsumura, Yukiko Ohyama, Tamotsu Kiyoshima, Seiji Nakamura, The diagnostic utility of submandibular gland sonography and labial salivary gland biopsy in IgG4-related dacryoadenitis and sialadenitis: Its potential application to the diagnostic criteria, Mod Rheumatol, 10.1080/14397595.2019.1576271, 30, 2, 379-384, 2020.03, Objectives: In this study, we investigated the diagnostic utility of submandibular gland (SMG) sonography and labial salivary gland (LSG) biopsy as a less invasive procedure for diagnosing IgG4-related dacryoadenitis and sialadenitis (IgG4-DS)Methods: Sixty-eight patients with suspected IgG4-DS by presenting swelling of elevated serum IgG (>1747 mg/dl) and/or swelling glands underwent SMG sonography, LSG biopsy and measurement for serum IgG4. SMG sonographic diagnosis was determined by the following characteristic changes; 'hypoechoic areas of a nodal pattern with high vascularity' and/or 'hypoechoic areas of a reticular pattern in the superficial part'.Results: Thirty-one patients were diagnosed with IgG4-DS, 5 with IgG4-RD unaccompanied by lacrimal and salivary gland lesions, 28 with Sjögren's syndrome, and 4 with malignant lymphoma. The sensitivity, specificity, and accuracy of SMG sonography and LSG biopsy were 100%, 83.8%, 91.2% and 64.5%, 73.8%, 75.0%, respectively. Moreover, those of SMG sonography and LSG biopsy combined with serum IgG4 concentration (>135 mg/dl) were 100%, 94.6%, 97.1% and 64.5%, 91.9%, 79.4%, respectively.Conclusion: LSG biopsy needs to be extremely careful to diagnose IgG4-DS because of its low sensitivity. SMG sonography is sufficient for the diagnosis of IgG4-DS, especially when combined with serologic analysis. Thus, SMG sonography could adapt to the diagnostic criteria of IgG4-DS as a non-invasive method..
9. Shinsuke Fujii, Yudai Tajiri, Kana Hasegawa, Shinji Matsumoto, Reiko U. Yoshimoto, Hiroko Wada, Shosei Kishida, Mizuho A. Kido, Hiromasa Yoshikawa, Satoru Ozeki, Tamotsu Kiyoshima, The TRPV4-AKT axis promotes oral squamous cell carcinoma cell proliferation via CaMKII activation., Laboratory Investigation, 10.1038/s41374-019-0357-z., 100, 2, 311-323, 2020.02, Most human malignant tumor cells arise from epithelial tissues, which show distinctive characteristics, such as polarization, cell-to-cell contact between neighboring cells, and anchoring to a basement membrane. When tumor cells invaginate into the stroma, the cells are exposed to extracellular environments, including the extracellular matrix (ECM). Increased ECM stiffness has been reported to promote cellular biological activities, such as excessive cellular growth and enhanced migration capability. Therefore, tumorous ECM stiffness is not only an important clinical tumor feature but also plays a pivotal role in tumor cell behavior. Transient receptor potential vanilloid 4 (TRPV4), a Ca2+-permeable nonselective cation channel, has been reported to be mechano-sensitive and to regulate tumorigenesis, but the underlying molecular mechanism in tumorigenesis remains unclear. The function of TRPV4 in oral squamous cell carcinoma (OSCC) is also unknown. The current study was conducted to investigate whether or not TRPV4 might be involved in OSCC tumorigenesis. TRPV4 mRNA levels were elevated in OSCC cell lines compared with normal oral epithelial cells, and its expression was required for TRPV4 agonist-dependent Ca2+ entry. TRPV4-depleted tumor cells exhibited decreased proliferation capabilities in three-dimensional culture but not in a low-attachment plastic dish. A xenograft tumor model demonstrated that TRPV4 expression was involved in cancer cell proliferation in vivo. Furthermore, loss-of-function experiments using siRNA or an inhibitor revealed that the TRPV4 expression was required for CaMKII-mediated AKT activation. Immunohistochemical analyses of tissue specimens obtained from 36 OSCC patients showed that TRPV4 was weakly observed in non-tumor regions but was strongly expressed in tumor lesions at high frequencies where phosphorylated AKT expression was frequently detected. These results suggest that the TRPV4/CaMKII/AKT axis, which might be activated by extracellular environments, promotes OSCC tumor cell growth..
10. Tanaka S, Kawanoa S, Hattori T, Matsubara R, Sakamoto T, Hashiguchi Y, Kanekoa N, Mikami Y, Morioka N, Maruse Y, Kitamura R, Hamada E, Hiwatashi M, Oobu K, Kiyoshima T, Nakamura S., Cytokeratin 19 as a biomarker of highly invasive oral squamous cell carcinoma with metastatic potential, Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology, 10.1016/j.ajoms.2019.10.007, 32, 1, 1-7, 2020.01, Objective
Cytokeratin (CK) 19 is a member of the acidic type I CK family. Recently, CK19 expression has been found in various tumor tissues; however, the significance of this remains unknown. The purpose of this study is to clarify the roles of CK19 in the progression of oral squamous cell carcinoma (OSCC).

Methods
A total of 100 patients who had been diagnosed with OSCC at our department between January 2011 and December 2016 was included. The patients were divided into three groups based on an optimal cut-off points (5% and 77%) of the labeling index (LI) as follows: group A; LI < 5%, group B; 5%≤ LI < 77%, group C; LI ≥ 77%. Then, clinicopathological features and survival rates were compared among the groups.

Results
Histologically high-grade tumors were significantly more common in group C than in groups A and B. Furthermore, the incidence of nodal metastasis was significantly higher in group C than in other groups. Intense CK19 immunoreactivity was detected in metastatic lymph nodes of groups B and C, but not from group A. Moreover, patients with advanced pN stage and extranodal extension were more common in groups B and C than group A. Disease-specific survival curves revealed poorer prognoses in group C.

Conclusions
These results suggest that CK19 is involved in OSCC invasion and metastasis and could be a novel biomarker of highly invasive OSCC with metastatic potential..
11. Noriko Ishiguro, Masafumi Moriyama, Katsuhiro Furusho, Sachiko Furukawa, Takuma Shibata, Yusuke Murakami, Akira Chinju, A. S. M. Rafiul Haque, Yuka Gion, Miho Ohta, Takashi Maehara, Akihiko Tanaka, Masaki Yamauchi, Mizuki Sakamoto, Keita Mochizuki, Yuko Ono, Jun‐Nosuke Hayashida, Yasuharu Sato, Tamotsu Kiyoshima, Hidetaka Yamamoto, Kensuke Miyake, Seiji Nakamura, Activated M2 Macrophages Contribute to the Pathogenesis of IgG4-Related Disease via Toll-Like Receptor 7/Interleukin-33 Signaling, ARTHRITIS & RHEUMATOLOGY, 10.1002/art.41052, 72, 1, 166-178, 2020.01, OBJECTIVE: IgG4-related disease (IgG4-RD) is a unique inflammatory disorder in which Th2 cytokines promote IgG4 production. In addition, recent studies have implicated the Toll-like receptor (TLR) pathway. This study was undertaken to examine the expression of TLRs in salivary glands (SGs) from patients with IgG4-RD.
METHODS: SGs from 15 patients with IgG4-RD, 15 patients with Sjögren's syndrome (SS), 10 patients with chronic sialadenitis, and 10 healthy controls were examined histologically. TLR family gene expression (TLR-1 through TLR-10) was analyzed by DNA microarray in the submandibular glands (SMGs). Up-regulation of TLRs was confirmed in SGs from patients with IgG4-RD. Finally, the phenotype of human TLR-7 (huTLR-7)-transgenic C57BL/6 mice was assessed before and after stimulation with TLR agonist.
RESULTS: In patients with IgG4-RD, TLR-4, TLR-7, TLR-8, and TLR-9 were overexpressed. Polymerase chain reaction validated the up-regulation of TLR-7 in IgG4-RD compared with the other groups. Immunohistochemical analysis confirmed strong infiltration of TLR-7-positive cells in the SGs of patients with IgG4-RD. Double immunohistochemical staining showed that TLR-7 expression colocalized with CD163+ M2 macrophages. After in vitro stimulation with a TLR-7 agonist, CD163+ M2 macrophages produced higher levels of interleukin-33 (IL-33), which is a Th2-activating cytokine. In huTLR-7-transgenic mice, the focus and fibrosis scores in SMGs, pancreas, and lungs were significantly higher than those in wild-type mice (P < 0.05). Moreover, the concentration of serum IgG, IgG1, and IL-33 in huTLR-7-transgenic mice was distinctly increased upon stimulation with a TLR-7 agonist (P < 0.05)..
12. Hisato Yoshida, Hitoshi Yoshimura, Shinpei Matsuda, Satoshi Yamamoto, Masahiro Ohmori, Minekatsu Taga, Keiichi Ohta, Takashi Ryoke, Hayato Itoi, Tamotsu Kiyoshima, Motohiro Kobayashi, and Kazuo Sano, Celecoxib suppresses lipopolysaccharide-stimulated oral squamous cell carcinoma proliferation in vitro and in vivo., Oncology letters., 10.3892/ol.2019.10975., 18, 6, 5793-5800, 2019.12, Periodontitis is one of the most common chronic oral inflammatory conditions worldwide and is associated with a risk of developing oral squamous cell carcinoma (OSCC). Porphyromonas gingivalis is a major pathogen in periodontitis, and its lipopolysaccharide (LPS) promotes the expression of cyclooxygenase-2 (COX-2) in OSCC both in vivo and in vitro. Celecoxib is a selective COX-2 inhibitor; however, its antitumor effects on P. gingivalis LPS-stimulated OSCC and the underlying molecular mechanism remain unclear. To elucidate the association between periodontitis and OSCC, the effect of P. gingivalis-derived LPS on OSCC cell proliferation was examined both in vitro and in vivo in the present study. The expression levels of COX-2 and p53 in OSCC cells with/without celecoxib treatment were determined via western blotting. The therapeutic potential of celecoxib in LPS-stimulated OSCC was evaluated by staining for Ki-67 and p21, as well as with terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling staining. LPS treatment significantly increased OSCC cell proliferation in vitro, and celecoxib significantly inhibited cell proliferation with/without LPS treatment. Celecoxib treatment of OSCC cells downregulated the protein expression levels of COX-2 compared with untreated cells, but there was little change in p53 expression. In the mouse xenograft model, oral administration of celecoxib significantly suppressed tumor growth, reduced the expression of Ki-67, increased the apoptosis index and induced p21 expression with/without LPS treatment. The results from the present study demonstrate that P. gingivalis' LPS can stimulate tumor growth by interacting with OSCC cells. In conclusion, these results suggest that celecoxib could be used for the effective prevention and treatment of LPS-stimulated OSCC..
13. A. S. M. Rafiul Haque, Masafumi Moriyama, Keigo Kubota, Noriko Ishiguro, Mizuki Sakamoto, Akira Chinju, Keita Mochizuki, Taiki Sakamoto, Naoki Kaneko, Ryusuke Munemura, Takashi Maehara, Akihiko Tanaka, Jun-Nosuke Hayashida, Shintaro Kawano, Tamotsu Kiyoshima, Seiji Nakamura, CD206(+) tumor-associated macrophages promote proliferation and invasion in oral squamous cell carcinoma via EGF production, Scientific Reports., 10.1038/s41598-019-51149-1, 9, 2019.10, Odontomas, developmental anomalies of tooth germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in tooth germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and tooth germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and tooth germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation..
14. Maehara T., Murakami Y., Kawano S., Mikami Y., Kiyoshima T., Chikui T., Kakizoe N., Munemura R., Nakamura S., Osteoid osteoma of mandibular bone: Case report and review of the literature, Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology, 10.1016/j.ajoms.2019.04.002, 31, 322-326, 322-326, 2019.09, Osteoid osteoma is a benign bone-forming tumor and characterized by its limited growth potential, not exceeding 2 cm. The radiological hallmark of this tumor is a nidus, which is a small round area of relative radiolucency. Osteoid osteoma can involve any bone but is most commonly found in long bones and is extremely rare in the head and neck region. This disease characteristically presents with dull pain, worse at night, and sometimes relieved with NSAIDs. A 24-year-old Japanese woman presented with spontaneous pain and tenderness on the lingual side of her mandibular second molar on the right side. The patient reported that her pain had gradually increased, becoming more continuous and severe and no longer responding to NSAIDs. An initial panoramic radiograph revealed an oval, internally non-uniform, somewhat obscure boundaries in the right mandible. Computed tomography (CT) scan revealed a sclerotic lesion with a delineated central calcified nidus surrounded by a radiolucent band. The interior of the nidus was a non-uniform, irregularly shaped area of high absorption. The nidus was removed with intralesional curettage under general anesthesia. The histopathology of the specimen consisted of actively proliferating osteoblasts mixed with an interlacing network of immature bone and osteoid trabeculae. Immunohistochemistry revealed that hardly detected osteoblasts or fibrous stromal cells with intense nuclear immunoreactivity for p16 and/or murine double minute 2 (mdm2). We thus distinguished the tumor from Low-grade central osteosarcoma (LGCO) with immunohistochemical findings. The histopathological diagnosis was thus osteoid osteoma..
15. Shinpei Matsuda, Hitoshi Yoshimura, Hisato Yoshida, Minekatsu Taga, Yoshiaki Imamura, Tamotsu Kiyoshima, Kazuo Sano, Ossifying fibroma in the mandibular angle mimicking metastatic clear cell renal cell carcinoma: A case report., Medicine (Baltimore)., 10.1097/MD.0000000000016595., 98, 33, e16595, 2019.08, RATIONALE: Ossifying fibroma is benign fibro-osseous neoplasm. The authors report a case of ossifying fibroma in the mandibular angle suspected as metastasis of clear cell renal cell carcinoma.
PATIENT CONCERNS: A 74-year-old man presented to the primary hospital complaining of frequent urination. A tumor in the left kidney was detected via an abdominal computed tomography scan. The patient then visited the Department of Urology at our hospital.
DIAGNOSES: According to whole-body imaging examinations, the patient was suspected of having renal cancer with mandibular metastasis. Also, a cystic lesion of the maxilla was revealed.
INTERVENTIONS: Left nephrectomy was performed by urologists, and the patient was diagnosed with clear cell renal cell carcinoma of the left kidney. Approximately 1 month later, resection with a safety margin of the mandibular lesion and removal of the maxillary lesion were performed by oral and maxillofacial surgeons.
OUTCOMES: The patient was diagnosed with ossifying fibroma of the mandible and an odontogenic keratocyst of the maxilla via a histopathological examination. Eighteen months have passed since the operation without clinical and imaging findings associated with recurrence.
LESSONS: Ossifying fibroma in the mandibular angle of elderly patients is extremely rare. Surgeons should consider the possibility of metastasis when osteolytic lesions of the jaw are found in patients with cancer..
16. Hitoshi Yoshimura, Hisato Yoshida, Shinpei Matsuda. Takashi Ryoke, Keiichi Ohta, Masahiro Ohmori, Satoshi Yamamoto, Tamotsu Kiyoshima, Motohiro Kobayashi, Kazuo Sano, The therapeutic potential of epigallocatechin‑3‑gallate against human oral squamous cell carcinoma through inhibition of cell proliferation and induction of apoptosis: In vitro and in vivo murine xenograft study., Molecular medicine reports., 10.3892/mmr.2019.10331, 2019.06, Oral squamous cell carcinoma (OSCC) is one of the most common malignant tumors in the oral region. Despite current therapeutic strategies, the survival rate has not been improved for several decades. Thus, it is important to develop a novel approach for the treatment of OSCC. Epigallocatechin‑3‑gallate (EGCG) is a major constituent of green tea and has previously been demonstrated to inhibit the growth of several types of cancer cells. However, few studies have investigated the effect of EGCG on human OSCC cells, especially in experimental animal models. The aim of the present study was to evaluate the therapeutic potential of EGCG for targeting human OSCC in vitro and in vivo. In the in vitro experiments, EGCG suppressed HSC‑3 cell viability in a time‑ and dose‑dependent manner. Cell cycle analysis revealed that EGCG induced G1 phase arrest of the tumor cells. Apoptosis was examined by Annexin V and propidium iodide staining, assays of caspase‑3 and -7 activity and TdT‑mediated dUTP nick end labeling (TUNEL) staining. Treatment with EGCG significantly increased caspase‑3 and -7 activities, and the percentage of apoptotic cells when compared with control cells. In the in vivo xenograft experiment on mice, EGCG treatment resulted in a 45.2% reduction in tumor size as compared with the control group without weight loss. In vivo cell proliferation and apoptosis were assessed by immunohistochemical Ki‑67 staining and the TUNEL staining. There were significant differences in Ki‑67 expression between the EGCG treatment group and control group, and the percentage of apoptotic cells in the EGCG treatment group was significantly greater than that in the control group. These results indicated that EGCG significantly inhibited cell proliferation by affecting the cell cycle progression and apoptosis in vitro and in vivo. These findings suggest that EGCG may have clinical applications as a novel approach to oral‑cancer therapy..
17. Reiko U. Yoshimoto, Reona Aijima, Yukiko Ohyama, Junko Yoshizumi, Tomoko Kitsuki, Yasuyoshi Ohsaki, Ai-Lin Cao, Atsushi Danjo, Yoshio Yamashita, Tamotsu Kiyoshima, Mizuho A. Kido, Impaired Junctions and Invaded Macrophages in Oral Epithelia With Oral Pain., Journal of Histochemistry & Cytochemistry, 10.1369/0022155418812405, 67, 4, 245-256, 2019.04, Recurrent or chronic oral pain is a great burden for patients. Recently, the links between epithelial barrier loss and disease were extended to include initiation and propagation. To explore the effects of pathohistological changes in oral epithelia on pain, we utilized labial mucosa samples in diagnostic labial gland biopsies from patients with suspected Sjögren's syndrome (SS), because they frequently experience pain and discomfort. In most labial mucosa samples from patients diagnosed with SS, disseminated epithelial cellular edema was prevalent as ballooning degeneration. The disrupted epithelia contained larger numbers of infiltrating macrophages in patients with oral pain than in patients without pain. Immunohistochemistry revealed that edematous areas were distinct from normal areas, with disarranged cell-cell adhesion molecules (filamentous actin, E-cadherin, β-catenin). Furthermore, edematous areas were devoid of immunostaining for transient receptor potential channel vanilloid 4 (TRPV4), a key molecule in adherens junctions. In an investigation on whether impaired TRPV4 affect cell-cell adhesion, calcium stimulation induced intimate cell-cell contacts among oral epithelial cells from wild-type mice, while intercellular spaces were apparent in cells from TRPV4-knockout mice. The present findings highlight the relationship between macrophages and epithelia in oral pain processing, and identify TRPV4-mediated cell-cell contacts as a possible target for pain treatment..
18. Shinsuke Fujii, Kengo Nagata, Shinji Matsumoto, Ken-ichi Kohashi, Akira Kikuchi, Yoshinao Oda, Tamotsu Kiyoshima, Naohisa Wada, Wnt/β-catenin signaling, which is activated in odontomas, reduces Sema3A expression to regulate odontogenic epithelial cell proliferation and tooth germ development., Scientific Reports., 10.1038/s41598-019-39686-1, 9, 2019.03, Odontomas, developmental anomalies of tooth germ, frequently occur in familial adenomatous polyposis patients with activated Wnt/β-catenin signaling. However, roles of Wnt/β-catenin signaling in odontomas or odontogenic cells are unclear. Herein, we investigated β-catenin expression in odontomas and functions of Wnt/β-catenin signaling in tooth germ development. β-catenin frequently accumulated in nucleus and/or cellular cytoplasm of odontogenic epithelial cells in human odontoma specimens, immunohistochemically. Wnt/β-catenin signaling inhibited odontogenic epithelial cell proliferation in both cell line and tooth germ development, while inducing immature epithelial bud formation. We identified Semaphorin 3A (Sema3A) as a downstream molecule of Wnt/β-catenin signaling and showed that Wnt/β-catenin signaling-dependent reduction of Sema3A expression resulted in suppressed odontogenic epithelial cell proliferation. Sema3A expression is required in appropriate epithelial budding morphogenesis. These results suggest that Wnt/β-catenin signaling negatively regulates odontogenic epithelial cell proliferation and tooth germ development through decreased-Sema3A expression, and aberrant activation of Wnt/β-catenin signaling may associate with odontoma formation..
19. Junko Yoshizumi, Hiroko Wada, Mayumi Shimizu, Yasufumi Horinouchi, Tamotsu Kiyoshima, Tetsuro Ikebe, Akimitsu Hirakia, A rare case of cemento-osseous dysplasia arising from a dislocated impacted tooth in the maxillary sinus., Journal of Oral and Maxillofacial Surgery, Medicine and Pathology, 10.1016/j.ajoms.2018.08.002, 31, 2, 94-97, 2019.03, Fibro-osseous and osteochondromatous lesions are non-neoplastic lesions containing metaplastic bone and/or cementum. The World Health Organization classification (4th edition, 2017) has categorized these lesions into subgroups: ossifying fibroma, familial gigantiform cementoma, fibrous dysplasia, cemento-osseous dysplasia and osteochondroma. We report a highly unusual case of cemento-osseous dysplasia in a 32-year-old male complaining of discomfort of the left cheek. The radiographic examination revealed that a spherical radiopaque lesion, which involved a tooth, was located in the left maxillary sinus. A definitive diagnosis could not be made based only on the radiographic and clinical findings. An excisional biopsy was performed by opening the left maxillary sinus under the general anaesthesia. Histopathological examination revealed that the lesion consists of fibrous tissue, woven bone and masses of cementum-like material associated with a dislocated tooth. Together with the radiographic, clinical and histopathological findings, the lesion was diagnosed as cemento-osseous dysplasia. Differential diagnosis between fibro-osseous lesions and tumor is critical; the present case highlights the importance of correlating clinical, radiological and histopathological findings in order to correctly diagnose and manage oral and maxillofacial lesions..
20. Yuma Hashiguchi, Shintarou Kawano, Yuichi Goto, Kaori Yasuda, Naoki Kaneko, Taiki Sakamoto, Ryota Matsubara, Teppei Jinno, Yasuyuki Maruse, Hideaki Tanaka, Masahiko Morioka, Taichi Hattori, Shoichi Tanaka, Tamotsu Kiyoshima, Seiji Nakamura, Tumor-suppressive roles of ΔNp63β-miR-205 axis in epithelial-mesenchymal transition of oral squamous cell carcinoma via targeting ZEB1 and ZEB2, Journal of Cellular Physiology, 10.1002/jcp.26267, 233, 10, 6565-6577, 2018.10, We previously revealed that epithelial-to-mesenchymal transition (EMT) was mediated by ΔNp63β, a splicing variant of ΔNp63, in oral squamous cell carcinoma (OSCC). Recent studies have highlighted the involvement of microRNA (miRNA) in EMT of cancer cells, though the mechanism remains unclear. To identify miRNAs responsible for ΔNp63β-mediated EMT, miRNA microarray analyses were performed by ΔNp63β-overexpression in OSCC cells; SQUU-B, which lacks ΔNp63 expression and displays EMT phenotypes. miRNAs microarray analyses revealed miR-205 was the most up-regulated following ΔNp63β-overexpression. In OSCC cells, miR-205 expression was positively associated with ΔNp63 and negatively with zinc-finger E-box binding homeobox (ZEB) 1 and ZEB2, potential targets of miR-205. miR-205 overexpression by miR-205 mimic transfection into SQUU-B cells led to decreasing ZEB1, ZEB2, and mesenchymal markers, increasing epithelial markers, and reducing cell motilities, suggesting inhibition of EMT phenotype. Interestingly, the results opposite to this phenomenon were obtained by transfection of miR-205 inhibitor into OSCC cells, which express ΔNp63 and miR-205. Furthermore, target protector analyses revealed direct regulation by miR-205 of ZEB1 and ZEB2 expression. These results showed tumor-suppressive roles of ΔNp63β and miR-205 by inhibiting EMT thorough modulating ZEB1 and ZEB2 expression in OSCC..
21. Yurie Mikami, Shinsuke Fujii, Ken-Ichi Kohashi, Yuichi Yamada, Masafumi Moriyama, Shintaro Kawano, Seiji Nakamura, Yoshinao Oda, Tamotsu Kiyoshima, Low-grade myofibroblastic sarcoma arising in the tip of the tongue with intravascular invasion: A case report., Oncology letters., 10.3892/ol.2018.9115, 16, 3, 3889-3894, 2018.09, Low-grade myofibroblastic sarcoma (LGMS) is a rare intermediate tumor, which rarely metastasizes and has myofibroblastic differentiation in various sites. It is particularly associated with the tongue in the head and neck region. The lack of any pathological features means it is difficult to make a conclusive diagnosis of LGMS. The immunohistochemical features and genomic rearrangements, including SS18-SSXs and MYH9-USP6s and the genetic mutations of cancer-associated genes, including APC, CTNNB1, EGFR, KRAS, PIK3CA and p53 were examined in a case of LGMS arising in the tip of the tongue. Immunohistochemically, the tumor cells were positive for alpha-smooth muscle actin and vimentin, as in previous reports. They demonstrated neither genomic rearrangements nor point mutations of cancer-associated genes. Although several tumor cells demonstrated intravascular invasion, the MIB-l labeling index of the cells was the same as the original lesion. To the best of our knowledge, this is the first case report of LGMS arising in the tip of the tongue with intravascular invasion..
22. Hisato Yoshida, Hitoshi Yoshimura, Shinpei Matsuda, Takashi Ryoke, Tamotsu Kiyoshima, Motohiro Kobayashi, Kazuo Sano, Effects of peritumoral bevacizumab injection against oral squamous cell carcinoma in a nude mouse xenograft model
A preliminary study, Oncology Letters, 10.3892/ol.2018.8399, 15, 6, 8627-8634, 2018.06, Angiogenesis serves a crucial role in tumor growth. Vascular endothelial growth factor (VEGF) is a potent regulator of tumor angiogenesis and is highly expressed in oral squamous cell carcinoma (OSCC). Bevacizumab, which binds to VEGF-A, inhibits the biological activity of VEGF and is clinically administered by intravenous injection. As intravenous chemotherapy intensifies the side effects experienced by OSCC patients, an alternative treatment option is desirable, particularly for older patients with OSCC who present with systemic disease complications. Generally, local injections of antitumor agents enhance tumoricidal activity and decrease side effects. However, the antitumor effects of peritumoral bevacizumab injections in OSCC are not fully understood. Therefore, the present study examined the effects of peritumoral bevacizumab injections in an experimental nude mouse model of OSCC through immunohistochemical staining for cluster of differentiation (CD)31 and α-smooth muscle actin (α‑SMA) and apoptosis assays. It was identified that peritumoral injections of bevacizumab significantly inhibited tumor growth in OSCC xenografts compared with peritumoral saline injections or no treatment (controls), and it was also revealed that treatment with bevacizumab significantly reduced CD31- and α-SMA-positive microvessel density (P<0.01) and increased level of tumor cell apoptosis (P<0.01) compared with the controls. In conclusion, these results collectively support the experimental basis for the clinical development of peritumoral bevacizumab injections for the treatment of OSCC..
23. Y. Maruse, Shintarou Kawano, Teppei Jinno, Ryota Matsubara, Y. Goto, N. Kaneko, T. Sakamoto, Y. Hashiguchi, M. Moriyama, T. Toyoshima, R. Kitamura, H. Tanaka, K. Oobu, T. Kiyoshima, S. Nakamura, Significant association of increased PD-L1 and PD-1 expression with nodal metastasis and a poor prognosis in oral squamous cell carcinoma, International Journal of Oral and Maxillofacial Surgery, 10.1016/j.ijom.2018.01.004, 47, 7, 836-845, 2018.07, Programmed cell death ligand 1 (PD-L1) and its receptor PD-1 are immune checkpoint molecules that attenuate the immune response. Blockade of PD-L1 enhances the immune response in a variety of tumours and thus serves as an effective anti-cancer treatment. However, the biological and prognostic roles of PD-L1/PD-1 signalling in oral squamous cell carcinoma (OSCC) remain to be elucidated. The purpose of this study was to examine the correlation of PD-L1/PD-1 signalling with the prognosis of OSCC patients to assess its potential therapeutic relevance. The expression of PD-L1 and of PD-1 was determined immunohistochemically in 97 patients with OSCC and the association of this expression with clinicopathological characteristics was examined. Increased expression of PD-L1 was found in 64.9% of OSCC cases and increased expression of PD-1 was found in 61.9%. Univariate and multivariate analysis revealed that increased expression of PD-L1 and PD-1 positively correlated with cervical lymph node metastasis. The expression of CD25, an activated T-cell marker, was negatively correlated with the labelling index of PD-L1 and PD-1. Moreover, the patient group with PD-L1-positive and PD-1-positive expression showed a more unfavourable prognosis than the group with PD-L1-negative and PD-1-negative expression. These data suggest that increased PD-L1 and PD-1 expression is predictive of nodal metastasis and a poor prognosis and is possibly involved in cancer progression via attenuating the immune response..
24. Takashi Maehara, Hamid Mattoo, Vinay S Mahajan, Samuel JH Murphy, Grace J Yuen, Noriko Ishiguro, Miho Ohta, Masafumi Moriyama, Takako Saeki, Hidetaka Yamamoto, Masaki Yamauchi, Joe Daccache, Tamotsu Kiyoshima, Seiji Nakamura, John H Stone, Shiv Pillai, The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo., Life science alliance., 10.26508/lsa.201800050, 1, 1, 2018.01, Distinct T follicular helper (TFH) subsets that influence specific class-switching events are assumed to exist, but the accumulation of isotype-specific TFH subsets in secondary lymphoid organs (SLOs) and tertiary lymphoid organs has not been hitherto demonstrated. IL-4-expressing TFH cells are surprisingly sparse in human SLOs. In contrast, in IgG4-related disease (IgG4-RD), a disorder characterized by polarized Ig class switching, most TFH cells in tertiary and SLOs make IL-4. Human IL-4+ TFH cells do not express GATA-3 but express nuclear BATF, and the transcriptomes of IL-4-secreting TFH cells differ from both PD1hi TFH cells that do not secrete IL-4 and IL-4-secreting non-TFH cells. Unlike IgG4-RD, IL-4+ TFH cells are rarely found in tertiary lymphoid organs in Sjögren's syndrome, a disorder in which IgG4 is not elevated. The proportion of CD4+IL-4+BATF+ T cells and CD4+IL-4+CXCR5+ T cells in IgG4-RD tissues correlates tightly with tissue IgG4 plasma cell numbers and plasma IgG4 levels in patients but not with the total plasma levels of other isotypes. These data describe a disease-related TFH subpopulation in human tertiary lymphoid organs and SLOs that is linked to IgG4 class switching..
25. Kana Ishibashi, kotaro ishii, Goro Sugiyama, Yu Kamata, Azusa Suzuki, Kumamaru Wataru, Yukiko Ohyama, Hiroyuki Nakano, Tamotsu Kiyoshima, Tomoki Sumida, Tomohiro Yamada, Yoshihide Mori, Regulation of β-catenin phosphorylation by PR55β in adenoid cystic carcinoma, Cancer Genomics and Proteomics, 10.21873/cgp.20064, 15, 1, 53-60, 2018.01, Background/Aim: Adenoid cystic carcinoma (AdCC) is a rare cancer of the salivary gland with high risk of recurrence and metastasis. Wnt signalling is critical for determining tumor grade in AdCC, as it regulates invasion and migration. β-catenin dephosphorylation plays an important role in the Wnt pathway, but its underlying molecular mechanism remains unclear. Materials and Methods: Because the regulatory subunits of protein phosphatase 2A (PP2A) drive Wnt signalling via target molecules, including β-catenin, we used qRT-PCR and immunoblot analysis to investigate the expression of these subunits in an AdCC cell line (ACCS) and a more aggressive subline (ACCS-M). Results: PR55β was highly expressed in ACCS-M, suggesting its functional importance. In addition, PR55β expression was associated with tumor grade, with ACCS-M exhibiting higher PR55β levels. More importantly, knockdown of PR55β in ACCS-M cells significantly reduced invasiveness and metastatic ability. Furthermore, dephosphorylation and total levels of β-catenin were dependent on PR55β in ACCS-M. Finally, we confirmed a correlation between PR55β staining intensity and histopathological type in human AdCC tissues. Conclusion: Our study provides new insight into the interaction between PR55β and β-catenin and suggests that PR55β may be a target for the clinical treatment of AdCC..
26. Sachiko Furukawa, Kazunari Oobu, Masafumi Moriyama, Shintarou Kawano, Saori Sako, Jun Nosuke Hayashida, Ryota Matsubara, Ken Ichi Ogata, Tamotsu Kiyoshima, Seiji Nakamura, Oral methotrexate-related lymphoproliferative disease presenting with severe osteonecrosis of the Jaw
A case report and literature review, Internal Medicine, 10.2169/internalmedicine.8946-17, 57, 4, 575-581, 2018.01, Long-term methotrexate (MTX) treatment can cause MTX-related lymphoproliferative disorder (MTX-LPD). We experienced a case of MTX-LPD that was associated with severe osteonecrosis of the jaw mimicking medication-related osteonecrosis of the jaw. The patient was an 81-year-old woman with rheumatoid arthritis (RA) who was treated with MTX and bisphosphonate. After 7 years, she was referred to our department for the assessment of giant ulcer and exposure of the alveolar bone of the left maxilla. Histopathological and immunological analyses confirmed a diagnosis of MTX-LPD. At seven months after the cessation of MTX treatment, the ulcerative and necrotic lesions had markedly decreased in size. A 1-year follow-up examination showed no evidence of recurrence and good RA control..
27. Yumiko I. Matsuishi, Hiroki Kato, Keiji Masuda, Haruyoshi Yamaza, Yuta Hirofuji, Hiroshi Sato, Hiroko Wada, Tamotsu Kiyoshima, Kazuaki Nonaka, Accelerated dentinogenesis by inhibiting the mitochondrial fission factor, dynamin related protein 1, Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2017.12.026, 495, 2, 1655-1660, 2018.01, Undifferentiated odontogenic epithelium and dental papilla cells differentiate into ameloblasts and odontoblasts, respectively, both of which are essential for tooth development. These differentiation processes involve dramatic functional and morphological changes of the cells. For these changes to occur, activation of mitochondrial functions, including ATP production, is extremely important. In addition, these changes are closely related to mitochondrial fission and fusion, known as mitochondrial dynamics. However, few studies have focused on the role of mitochondrial dynamics in tooth development. The purpose of this study was to clarify this role. We used mouse tooth germ organ cultures and a mouse dental papilla cell line with the ability to differentiate into odontoblasts, in combination with knockdown of the mitochondrial fission factor, dynamin related protein (DRP)1. In organ cultures of the mouse first molar, tooth germ developed to the early bell stage. The amount of dentin formed under DRP1 inhibition was significantly larger than that of the control. In experiments using a mouse dental papilla cell line, differentiation into odontoblasts was enhanced by inhibiting DRP1. This was associated with increased mitochondrial elongation and ATP production compared to the control. These results suggest that DRP1 inhibition accelerates dentin formation through mitochondrial elongation and activation. This raises the possibility that DRP1 might be a therapeutic target for developmental disorders of teeth..
28. Naoki Kaneko, Shintarou Kawano, Kaori Yasuda, Yuma Hashiguchi, Taiki Sakamoto, Ryota Matsubara, Yuichi Goto, Teppei Jinno, Yasuyuki Maruse, Masahiko Morioka, Taichi Hattori, Shoichi Tanaka, Hideaki Tanaka, Tamotsu Kiyoshima, Seiji Nakamura, Differential roles of kallikrein-related peptidase 6 in malignant transformation and ΔNp63β-mediated epithelial-mesenchymal transition of oral squamous cell carcinoma, Oral Oncology, 10.1016/j.oraloncology.2017.11.004, 75, 148-157, 2017.12, We previously reported that epithelial-to-mesenchymal transition (EMT) was mediated by ΔNp63β in oral squamous cell carcinoma (OSCC). In this study, DNA microarray analyses were performed using ΔNp63β-overexpressing OSCC cells to identify genes associated with ΔNp63β-mediated EMT. Thereby, we focused on kallikrein-related peptidase (KLK) 6, most up-regulated following ΔNp63β-overexpression, that activates protease-activated receptors (PARs). In RT-PCR analyses, ΔNp63 was positively associated with KLK6 and PAR2 and negatively with PAR1 in OSCC cells. By ΔNp63 knockdown, KLK6 and PAR2 expression was decreased and PAR1 was increased. Furthermore, KLK6 knockdown led to enhancing migration and invasion, and inhibiting proliferation, suggesting EMT-phenotypes. Although, in the KLK6 or PAR2 knockdown cells, phosphorylation of ERK was reduced, it was restored in the KLK6 knockdown OSCC cells treated with recombinant KLK6 proteins. Immunohistochemistry showed ΔNp63, KLK6, and PAR2 were more strongly expressed in the epithelial dysplasia and central region of OSCC than normal oral epithelium, whereas PAR1 expression was undetectable. Interestingly, at the invasive front of OSCC, ΔNp63, KLK6, and PAR2 were reduced, but PAR1 was elevated. In addition, the OSCC patients with decreasing KLK6 expression at the invasive front had more unfavourable prognosis. These results suggested differential roles of KLK6 in malignant transformation and EMT; high ΔNp63β expression up-regulates KLK6-PAR2 and down-regulates PAR1, inducing malignant transformation in oral epithelium with stimulating proliferation through ERK signal activation. Moreover, KLK6-PAR2 expression is down-regulated and PAR1 is up-regulated when ΔNp63β expression is decreased, leading to EMT with enhancing migration and invasion through ERK signal reduction at the invasive front..
29. Taiki Sakamoto, Shintaro Kawano, Ryota Matsubara, Yuichi Goto, Teppei Jinno, Yasuyuki Maruse, Naoki Kaneko, Yuma Hashiguchi, Taichi Hattrori, Shoichi Tanaka, Ryoji Kitamura, Tamotsu Kiyoshima, Seiji Nakamura, Critical roles of Wnt5a-Ror2 signaling in aggressiveness of tongue squamous cell carcinoma and production of matrix metalloproteinase-2 via ΔNp63β-mediated epithelial-mesenchymal transition., Oral Oncology, 10.1016/j.oraloncology.2017.03.019, 69, 15-25, 2017.06, OBJECTIVES: We previously showed that ΔNp63β, a splicing variant of ΔNp63, mediated EMT and affected cell motility. DNA microarray was thus performed to elucidate the mechanism that ΔNp63β affects cell motility. As the results, Wnt5a was significantly down-regulated by ΔNp63β overexpression in tongue SCC cell line (SQUU-B) with EMT phenotype.
MATERIALS AND METHODS:
Seven OSCC cell lines were used. Expression of ΔNp63, Wnt5a, its receptor Ror2, and matrix metalloproteinases (MMPs) were analyzed by RT-PCR, real-time PCR, and western blotting, and gelatin zymography. Furthermore, we examined the effects of siRNA for Wnt5a or Ror2 and recombinant human Wnt5a (rhWnt5a) on motility of tongue SCC cells. Biopsy specimens from tongue SCC patients were used for immunohistochemical staining of Wnt5a and Ror2.
RESULTS:
Wnt5a and Ror2 were expressed only in SQUU-B cells without ΔNp63 expression, and negatively associated with ΔNp63 expression in other cells. ΔNp63β overexpression in SQUU-B cells decreased Wnt5a and Ror2 expression. By Wnt5a or Ror2 knockdown, cell motility was remarkably inhibited, but EMT markers expression was unaffected. MMP-2 expression and the activities inversely correlated with ΔNp63 expression, and were inhibited by Wnt5a or Ror2 knockdown. Cell motility and MMP-2 activities were recovered by adding rhWnt5a in the cells with Wnt5a knockdown, but not in those with Ror2 knockdown. Moreover, immunohistochemical analyses in tongue SCC specimens found that high expression of Wnt5a or Ror2 was associated with poorer prognosis.
CONCLUSION:
Wnt5a-Ror2 signaling enhanced tongue SCC cell aggressiveness and promoted production of MMP-2 following ΔNp63β-mediated EMT..
30. Keigo Kubota, Masafumi Moriyama, Sachiko Furukawa, HASM Rafiul, Yasuyuki Maruse, Teppei Jinno, Akihiko Tanaka, Miho Ohta, Noriko Ishiguro, Masaki Yamauchi, Mizuki Sakamoto, Takashi Maehara, Hayashida Jun-Nosuke, Shintaro Kawano, Tamotsu Kiyoshima, Seiji Nakamura, CD163(+) CD204(+) tumor-associated macrophages contribute to T cell regulation via interleukin-10 and PD-L1 production in oral squamous cell carcinoma, SCIENTIFIC REPORTS, 10.1038/s41598-017-01661-z, 7, 1, 1755, 2017.05, Tumor-associated macrophages (TAMs) promote cancer cell proliferation, invasion, and metastasis by producing various mediators. Although preclinical studies demonstrated that TAMs preferentially express CD163 and CD204, the TAM subsets in oral squamous cell carcinoma (OSCC) remain unknown. In this study, we examined the expression and role of TAM subsets in OSCC. Forty-six patients with OSCC were analyzed for expression of TAMs in biopsy samples by immunohistochemistry. We examined TAM subsets and their production of immune suppressive molecules (IL-10 and PD-L1) in peripheral blood mononuclear cells from three OSCC patients by flow cytometry. CD163 was detected around the tumor or connective tissue, while CD204 was detected in/around the tumors. Flow cytometric analysis revealed that CD163+CD204+ TAMs strongly produced IL-10 and PD-L1 in comparison with CD163+CD204- and CD163-CD204+ TAMs. Furthermore, the number of activated CD3+ T cells after co-culture with CD163+CD204+ TAMs was significantly lower than that after co-culture with other TAM subsets. In clinical findings, the number of CD163+CD204+ TAMs was negatively correlated with that of CD25+ cells and 5-year progression-free survival. These results suggest that CD163+CD204+ TAMs possibly play a key role in the invasion and metastasis of OSCC by T-cell regulation via IL-10 and PD-L1 production..
31. Eiji Mitate, Shintaro Kawano, Yurie Mikami, Tamotsu Kiyoshima, Tetsuro Ikebe, Seiji Nakamura, A case of inversely fused tooth of impacted maxillary third molar and supernumerary tooth, International Journal of Case Reports and Images, 10.5348/ijcri-201723-CR-10762, 8, 2, 129-132, 2017.03, We report an extremely rare case of an impacted maxillary third molar inversely fused with a supernumerary tooth in a 51-year-old male. The panoramic and dental radiographs show a radiopaque and tooth-like mass of 20×15 mm located in the third maxillary region of the right maxilla. Computed tomography revealed that the tooth-like structure was a union of the impacted maxillary third molar with a inversed supernumerary tooth, creating a fused tooth with a common dental pulp. To our knowledge, only one case of inversely fusion of a maxillary third molar with a supernumerary tooth has been reported..
32. Yurie Mikami, Shinsuke Fujii, Kengo Nagata, Hiroko Wada, Kana Hasegawa, Misaki Abe, Reiko U Yoshimoto, Shintaro Kawano, Seiji Nakamura, Tamotsu Kiyoshima, GLI-mediated Keratin 17 expression promotes tumor cell growth through the anti-apoptotic function in oral squamous cell carcinomas., J Cancer Res Clin Oncol, 10.1007/s00432-017-2398-2, 2017.03, PURPOSE:Keratin 17 (KRT17) has been suggested as a potential diagnostic marker of squamous cell carcinoma including oral squamous cell carcinoma (OSCC). The current study was conducted to clarify the function of KRT17 and its expression mechanism in OSCC.METHODS:Immunohistochemical analyses were carried out to examine the expression of KRT17, GLI family zinc finger (GLI)-1, GLI-2, or cleaved caspase-3 in OSCCs. The expression of KRT17, GLI-1, or GLI-2 was investigated among OSCC cell lines, and the effects of loss-of-function of KRT17 or GLI, using siRNA or inhibitor, on the cell growth of the OSCC cell line HSC-2 particularly with respect to apoptosis were examined.RESULTS:Immunohistochemical analyses of tissue specimens obtained from 78 OSCC patients revealed that KRT17 was not observed in non-tumor regions but was strongly expressed at high frequencies in tumor regions. Knockdown of KRT17 increased the number of cleaved caspase-3-positive cells, leading to the reduction of cell number. Loss-of-function of GLI-1 or GLI-2 also increased the cell numbers of apoptotic cells positive for staining of Annexin-V and propidium iodide (PI) and the terminal deoxynucleotidyl transferase dUTP-biotin nick-end labeling (TUNEL) method, and induced DNA fragmentation. This inhibitory effect on cell growth was partially rescued by exogenous KRT17 expression. In the KRT17-positive regions in OSCCs, GLI-1 or GLI-2 was frequently detected, and the number of cells with cleaved caspase-3 positive was decreased.CONCLUSIONS:KRT17 promotes tumor cell growth, at least partially, through its anti-apoptotic effect as a result of the KRT17 overexpression by GLIs in OSCC..
33. Masafumi Moriyama, Miho Ohta, Sachiko Furukawa, Yurie Mikami, Akihiko Tanaka, Takashi Maehara, Masaki Yamauchi, Noriko Ishiguro, Jun-Nosuke Hayashida, Shintaro Kawano, Yukiko Ohyama, Tamotsu Kiyoshima, Seiji Nakamura, The diagnostic utility of labial salivary gland biopsy in IgG4-related disease., Mod Rheumatol., 10.3109/14397595.2016.1148225., 26, 5, 725-729, 2016.09, Objective: For the definitive diagnosis of IgG4-related disease (IgG4-RD), biopsies of local lesions are recommended so as to exclude other diseases, including lymphoma and cancer. However, performing biopsies of underlying organs is technically difficult. In this study, we examined the diagnostic utility of labial salivary gland (LSG) biopsy as a less invasive procedure.
METHODS: Sixty-six patients with suspected IgG4-RD by clinical findings or high serum IgG4 underwent LSG biopsy. We examined the relationship between the number of IgG4-positive plasma cells in LSG and clinical findings.
RESULTS: The final diagnosis was 45 patients with IgG4-RD, 12 with Sjögren's syndrome, four with suspected Sjögren's syndrome, three with malignant lymphoma, one with systemic lupus erythematosus, and one with Warthin's tumor. The sensitivity, specificity, and accuracy of LSG biopsy were 55.6%, 100.0%, and 70.0%, respectively. Forty-five IgG4-RD patients were divided into two groups: 1) 25 with lesions of salivary glands (IgG4-RD S+) and 2) 20 without these lesions (IgG4-RD S-). Seventeen of 25 (68.0%) IgG4-RD S + and 8 of 20 (40.0%) IgG4-RD S - patients were positive for LSG biopsy. In the IgG4-RD S - patients, the mean number of affected organs and serum IgG4 in the positive cases for LSG biopsy were significantly higher than in the negative cases.
CONCLUSION: A solo LSG biopsy is insufficient for the diagnosis of IgG4-RD because of its low sensitivity. However, LSG biopsy combined with clinical findings, including serum IgG4 and number of affected organs, may contribute towards a diagnosis of IgG4-RD patients with affected underlying organs..
34. Kana Hasegawa, Hiroko Wada, Kengo Nagata, Hiroaki Fujiwara, Naohisa Wada, Hirotaka Someya, Yurie Mikami, Hidetaka Sakai, Tamotsu Kiyoshima, Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse tooth germ development, Int. J. Mol. Med., 10.1007/s10735-016-9680-5, 47, 4, 375-387, 2016.08.
35. Shintaro Kawano, Y Zeng, Kazunari Oobu, Ryota Matsubara, Yuichi Goto, Toru Chikui, Tadamasa Yoshitake, Tamotsu Kiyoshima, Teppei Jinno, Yasuyuki Maruse, Eiji Mitate, Ryoji Kitamura, Hideaki Tanaka, Takeshi Toyoshima, Tsuyoshi Sugiura, Seiji Nakamura, Clinicopathological evaluation of pre-operative chemoradiotherapy with S-1 as a treatment for locally advanced oral squamous cell carcinoma, Oncol Lett., 10.3892/ol.2016.4411, 11, 5, 3369-3376, 2016.05, The administration of pre-operative chemotherapy with S-1 and concurrent radiotherapy at a total dose of 30 Gy was clinicopathologically evaluated as a treatment for locally advanced oral squamous cell carcinoma (OSCC) in the present study. The participants comprised 81 patients with OSCC, consisting of 29 patients with stage II disease, 12 patients with stage III disease and 40 patients with stage IV disease. All patients received a total radiation dose of 30 Gy in daily fractions of 2 Gy, 5 times a week, for 3 weeks, and the patients were concurrently administered S-1 at a dose of 80-120 mg, twice daily, over 4 consecutive weeks. Radical surgery was performed in all cases at 2-6 weeks subsequent to the end of pre-operative chemoradiotherapy. The most common adverse event was oropharyngeal mucositis, but this was transient in all patients. No severe hematological or non-hematological toxicities were observed. The clinical and histopathological response rates were 70.4 and 75.3%, respectively. Post-operatively, local failure developed in 6 patients (7.4%) and neck failure developed in 2 patients (2.5%). Distant metastases were found in 7 patients (8.6%). The overall survival rate, disease-specific survival rate and locoregional control rate at 5 years were 87.7, 89.9 and 90.6%, respectively. Locoregional recurrence occurred more frequently in patients that demonstrated a poor histopathological response compared with patients that demonstrated a good response (P<0.01). These results indicate that pre-operative S-1 chemotherapy with radiotherapy at a total dose of 30 Gy is feasible and effective for patients with locally advanced OSCC, and that little or no histopathological response may be a risk factor for locoregional recurrence in this treatment..
36. Rumi Yoshihama, Koujiro Yamaguchi, Tsuyoshi Sugiura, Ikumi Imajyo, Mariko Mine, Naomi Hiyake, Naonari Akimoto, Yosuke Kobayashi, Satomi Chigita, Kumamaru Wataru, Tamotsu Kiyoshima, Yoshihide Mori, Expression of SOX2, KLF4, and Brachyury transcription factor is correlated with metastasis and poor prognosis in oral squamous cell carcinoma patients., Oncol Lett. , 11, 2, 1435-1446, 2016.02.
37. Miho Ohta, Masafumi Moriyama, Takashi Maehara, Yuka Gion, Sachiko Furukawa, Akihiko Tanaka, Hayashida Jun-Nosuke, Masaki Yamauchi, Noriko Ishiguro, Yurie Mikami, Hiroto Tsuboi, Mana Iizuka-Koga, Shintaro Kawano, Yasuharu Sato, Tamotsu Kiyoshima, Takayuki Sumida, Seiji Nakamura, DNA Microarray Analysis of Submandibular Glands in IgG4-Related Disease Indicates a Role for MARCO and Other Innate Immune-Related Proteins., Medicine (Baltimore), doi: 10.1097/MD.0000000000002853., 95, 7, e2853, 2016.02.
38. Yasuyuki Maruse, Shintaro Kawano, Tamotsu Kiyoshima, Yuichi Goto, Ryota Matsubara, Toru Chikui, Daigo Yoshiga, Seiji Nakamura, Case of mucoepidermoid carcinoma of the sublingual gland accompanied with extensive dystrophic calcification and intratumoral bone formation, Head Neck., 10.1002/hed.24036, 37, 11, E161-E164, 2015.11.
39. Kenji Ueki, Yuta Matsukuma, Masutani K, Akihiko Tsuchimoto, Kiichiro Fujisaki, Kumiko Torisu, Shigeru Tanaka, Tamotsu Kiyoshima, Satoshi Hisano, Kitazono T, Kazuhiko Tsuruya, Membranoproliferative glomerulonephritis with predominant IgG2 and IgG3 deposition in a patient with IgG4-related disease, BMC NEPHROLOGY, 10.1186/s12882-015-0164-8, 16, 2015.10.
40. KEIJI MASUDA, Shintaro Kawano, Haruyoshi Yamaza, Taiki Sakamoto, Tamotsu Kiyoshima, Seiji Nakamura, Kazuaki Nonaka, Complete resolution of a calcifying cystic odontogenic tumor with physiological eruption of a dislocated permanent tooth after marsupialization in a child with a mixed dentition: a case report, WORLD JOURNAL OF SURGICAL ONCOLOGY, 10.1186/s12957-015-0697-0, 13, 2015.09.
41. Yoshikazu Hayashi, Masafumi Moriyama, Takashi Maehara, Yuichi Goto, Shintaro Kawano, Miho Ohta, Akihiko Tanaka, Shoko Furukawa, Hayashida Jun-Nosuke, Tamotsu Kiyoshima, Mayumi Shimizu, Toru Chikui, Seiji Nakamura, A case of mantle cell lymphoma presenting as IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease., World J Surg Oncol., doi: 10.1186/s12957-015-0644-0., 13, 225, 2015.07.
42. Teppei Jinno, Shintaro Kawano, Yasuyuki Maruse, Ryota Matsubara, Yuichi Goto, Taiki Sakamoto, Yuma Hashiguchi, Naoki Kaneko, Hideaki Tanaka, Ryoji Kitamura, Takeshi Toyoshima, Akiko Jinno, Masafumi Moriyama, Kazunari Oobu, Tamotsu Kiyoshima, Seiji Nakamura, Increased expression of interleukin-6 predicts poor response to chemoradiotherapy and unfavorable prognosis in oral squamous cell carcinoma., Oncol Rep., doi: 10.3892/or.2015.3838., 33, 5, 2161-2168, 2015.05.
43. Hirotaka Someya, Fujiwara Hiroaki, Kengo Nagata, Hiroko Wada, Kana Hasegawa, Yurie Mikami, Akiko Jinno, Hidetaka Sakai, Kiyoshi Koyano, Tamotsu Kiyoshima, Thymosin beta 4 is associated with RUNX2 expression via the Smad and Akt signaling pathways in mouse dental epithelial cells., Int. J. Mol. Med., doi: 10.3892/ijmm.2015.2118., 35, 5, 1169-1178, 2015.05.
44. Takashi Maehara, Masafumi Moriyama, Shintaro Kawano, Hayashida Jun-Nosuke, Shoko Furukawa, Miho Ohta, Akihiko Tanaka, Masaki Yamauchi, YUKIKO OHYAMA, Tamotsu Kiyoshima, Seiji Nakamura, Cytokine profiles contribute to understanding the pathogenic difference between good syndrome and oral lichen planus: two case reports and literature review.
, Medicine (Baltimore), doi: 10.1097/MD.0000000000000704., 94, 14, e704, 2015.04.
45. Sachiko Furukawa, Masafumi Moriyama, Shintaro Kawano, Akihiko Tanaka, Takashi Maehara, Hayashida Jun-Nosuke, Yuichi Goto, Tamotsu Kiyoshima, Hideki Shiratsuchi, YUKIKO OHYAMA, Miho Ohta, Yumi Imabayashi, Seiji Nakamura, Clinical relevance of Küttner tumour and IgG4-related dacryoadenitis and sialoadenitis., Oral diseases, 10.1111/odi.12259. , 21, 2, 257-262, 2015.03.
46. Miho Ohta, Masafumi Moriyama, Yuichi Goto, Shintaro Kawano, Akihiko Tanaka, Takashi Maehara, Shoko Furukawa, Hayashida Jun-Nosuke, Tamotsu Kiyoshima, Mayumi Shimizu, Yojiro Arinobu, Seiji Nakamura, A case of marginal zone B cell lymphoma mimicking IgG4-related dacryoadenitis and sialoadenitis., World J Surg Oncol., doi: 10.1186/s12957-015-0459-z., 13, 67, 2015.02.
47. Masafumi Moriyama, Shoko Furukawa, Shintaro Kawano, Yuichi Goto, Tamotsu Kiyoshima, Akihiko Tanaka, Takashi Maehara, Hayashida Jun-Nosuke, Miho Ohta, Seiji Nakamura, The diagnostic utility of biopsies from the submandibular and labial salivary glands in IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease., Int J Oral Maxillofac Surg., doi: 10.1016/j.ijom.2014.06.014., 43, 10, 1276-1281, 2014.10.
48. Makiko Kihara, Tamotsu Kiyoshima, Kengo Nagata, Hiroko Wada, Fujiwara Hiroaki, Kana Hasegawa, Hirotaka Someya, Ichiro Takahashi, Hidetaka Sakai, Itm2a expression in the developing mouse first lower molar, and the subcellular localization of itm2a in mouse dental epithelial cells., PLoS One., doi: 10.1371/journal.pone.0103928., 9, 7, e103928, 2014.07.
49. Goro Sugiyama, YUKIKO OHYAMA, MASANORI SASAKI, Tamotsu Kiyoshima, Mayumi Shimizu, Kaneki Eisuke, Yasuharu TAKENOSHITA, Metastatic adenocarcinoma of the mandibular condyle from uterine cervix: Report of a case., Oral Sci Int., 11, 1, 40-44, 2014.01.
50. Tamotsu Kiyoshima, Fujiwara Hiroaki, Kengo Nagata, Hiroko Wada, Yukiko Okuma, Maho Shiotsuka, Makiko Kihara, Kana Hasegawa, Hirotaka Someya, Hidetaka Sakai, Induction of dental epithelial cell differentiation marker gene expression in non-odontogenic human keratinocytes by transfection with thymosin beta 4., Stem Cell Research, doi: 10.1016/j.scr.2013.11.006., 12, 1, 309-322, 2014.01.
51. Maho Shiotsuka, Hiroko Wada, Tamotsu Kiyoshima, Kengo Nagata, Fujiwara Hiroaki, Makiko Kihara, Kana Hasegawa, Hirotaka Someya, Ichiro Takahashi, Hidetaka Sakai, The expression and function of thymosin beta 10 in tooth germ development., Int J Dev Biol, doi: 10.1387/ijdb.120240hs., 57, 11-12, 873-883, 2013.12.
52. Tamotsu Kiyoshima, Hisato Yoshida, Hiroko Wada, Kengo Nagata, Fujiwara Hiroaki, Makiko Kihara, Kana Hasegawa, Hirotaka Someya, Hidetaka Sakai, Chemoresistance to Concanamycin A1 in Human Oral Squamous Cell Carcinoma Is Attenuated by an HDAC Inhibitor Partly via Suppression of Bcl-2 Expression, PLOS ONE, 10.1371/journal.pone.0080998, 8, 11, 2013.11.
53. Eiji Mitate, Shintaro Kawano, Tamotsu Kiyoshima, Toshiyuki Kawazu, Toru Chikui, Yuichi Goto, Ryota Matsubara, Seiji Nakamura, Carcinoma ex pleomorphic adenoma of the upper lip: a case of an unusual malignant component of squamous cell carcinoma, WORLD JOURNAL OF SURGICAL ONCOLOGY, 10.1186/1477-7819-11-234, 11, 2013.09.
54. Tamotsu Kiyoshima, Kengo Nagata, Hiroko Wada, Fujiwara Hiroaki, Maho Shiotsuka, Makiko Kihara, Kana Hasegawa, Hirotaka Someya, Hidetaka Sakai, Immunohistochemical Expression of Thymosin β4 in Ameloblastomas and Odontomas., Histol. Histopathol., doi: 10.3892/ijo.2012.1594., 28, 6, 775-786, 2013.06.
55. Toyoshima Takeshi, Hideaki Tanaka, Ryoji Kitamura, Tamotsu Kiyoshima, Yuji SHIRATSUCHI, Seiji Nakamura, Traumatic ciliated cyst derived from zygomaticomaxillary fracture: Report of a case, J. Oral Maxillofac. Surg. Med. Pathol., doi.org/10.1016/j.ajoms.2013.01.004, in press, 2013.03.
56. Yukiko Okuma, Tamotsu Kiyoshima, Ieyoshi Kobayashi, Kengo Nagata, Hiroko Wada, Fujiwara Hiroaki, Haruyoshi Yamaza, Kazuaki Nonaka, Hidetaka Sakai, Multiple functional involvement of Thymosin beta-4 in tooth germ development, HISTOCHEMISTRY AND CELL BIOLOGY, 10.1007/s00418-012-1033-1, 139, 2, 355-370, 2013.02.
57. Tamotsu Kiyoshima, Norio Enoki, Ieyoshi Kobayashi, Takako Sakai, Kengo Nagata, Hiroko Wada, Fujiwara Hiroaki, 大隈 由紀子, Hidetaka Sakai, Oxidative stress caused by a low concentration of hydrogen peroxide induces senescence-like changes in mouse gingival fibroblasts., Int. J. Mol. Med., doi: 10.3892/ijmm.2012.1102., 30, 5, 1007-1012, 2012.11.
58. Lutfun Naher, Tamotsu Kiyoshima, Ieyoshi Kobayashi, Hiroko Wada, Kengo Nagata, Fujiwara Hiroaki, 大隈 由紀子, Satoru Ozeki, Seiji Nakamura, Hidetaka Sakai, STAT3 signal transduction through interleukin-22 in oral squamous cell carcinoma., Int. J. Oncol., doi: 10.3892/ijo.2012.1594., 41, 5, 1577-1586, 2012.11.
59. Hiroe Kakehashi, Shintaro Kawano, Tamotsu Kiyoshima, Mayumi Shimizu, Masafumi Moriyama, Ryota Matsubara, Takahiro Kiyosue, Yuichi Goto, Hideki Shiratsuchi, Seiji Nakamura, Parotid gland myoepithelioma with remarkable cystic formation: A case report, Journal of Oral and Maxillofacial Surgery, Medicine and Pathology, in press, 2012.07.
60. Honda JY, Kobayashi I, Kiyoshima T, Nagata K, Wada H, Ookuma Y, Fujiwara H, Shiotsuka M, Takahashi I, Sakai H., In situ expression of the mitochondrial ATPase6 gene in the developing tooth germ of the mouse lower first molar., J Mol Histol., 42(1):83-90., 2011.02.
61. Takahashi KF, Kiyoshima T, Kobayashi I, Xie M, Yamaza H, Fujiwara H, Ookuma Y, Nagata K, Wada H, Sakai T, Terada Y, Sakai H., Protogenin, a new member of the immunoglobulin superfamily, is implicated in the development of the mouse lower first molar., BMC Dev Biol., 10:115-129, 2010.11.
62. Akhter M, Kobayashi I, Kiyoshima T, Nagata K, Wada H, Ookuma Y, Fujiwara H, Honda JY, Sakai H., In situ expression of 15 kDa interferon alpha responsive gene in the developing tooth germ of the mouse lower first molar., J Mol Histol., 41(4-5):185-91, 2010.10.
63. Xie M., Kobayashi I., Kiyoshima T., Nagata K., Ookuma Y., Fujiwara H. and Sakai H., In situ expression of ribosomal protein L21 in developing tooth germ of the mouse lower first molar. , J Mol Histol., 40:361-7., 2009.12.
64. Honda JY, Kobayashi I, Kiyoshima T, Yamaza H, Xie M, Takahashi K, Enoki N, Nagata K, Nakashima A, Sakai H., Glycolytic enzyme Pgk1 is strongly expressed in the developing tooth germ of the mouse lower first molar., J Mol Histol., 3(4):423-32., 2008.04.
65. Xie M, Kobayashi I, Kiyoshima T, Yamaza H, Honda JY, Takahashi K, Enoki N, Akamine A, Sakai H., Functional implication of nucleolin in the mouse first molar development., J Biol Chem., 282(32):23275-83. , 2007.08.
66. Enoki N, Kiyoshima T, Sakai T, Kobayashi I, Takahashi K, Terada Y, Sakai H., Age-dependent changes in cell proliferation and cell death in the periodontal tissue and the submandibular gland in mice: a comparison with other tissues and organs., J Mol Histol., 38(4):321-32., 2007.08.
67. Ieyoshi Kobayashi, Tamotsu Kiyoshima, Hiroko Wada, Kou Matsuo, Kazuaki Nonaka, Jun-ya Honda, Kiyoshi Koyano, Hidetaka Sakai, Type II/III Runx2/Cbfa1 is required for tooth germ development, Bone, 38: 836-844, 2006.06.
68. Hideaki Fukuzawa, Tamotsu Kiyoshima, Ieyoshi Kobayashi, Satoru Ozeki, Hidetaka Sakai, Transcription promoter activity of the human S100A7 gene in oral squamous cell carcinoma cell lines., Biochem, Biophys.Acta, 1759: 171-176,, 2006.04.
69. Ken-ichiro Hashimoto, Tamotsu Kiyoshima, Kou Matsuo, Satoru Ozeki, Hidetaka Sakai, Effect of SCCA1 and SCCA2 on the suppression of TNF-a-induced cell death by impeding the release of mitochondrial cytochrome c in oral squamous cell carcinoma cell line., Tumor Biol., 10.1159/000086949, 26, 4, 165-172, 26:165-172, 2005.07.
70. Merina Akhter, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Kou Matsuo, Haruyoshi Yamaza, Hiroko Wada, Jun-ya Honda, Xie Ming, Hidetaka Sakai, Possible functional involvement of thymosin beta 4 in developing tooth germ of mouse lower first molar., Histochem. Cell Biol., 10.1007/s00418-005-0040-x, 124, 3-4, 207-213, 124: 207-213, 2005.01.
71. Paz J, Wade K, Kiyoshima T, Sodek J, Tang J, Tu Q, Yamauchi M, Chen J., Tissue- and bone cell-specific expression of bone sialoprotein is directed by a 9.0 kb promoter in transgenic mice., Matrix Biol., 24(5):341-52, 2005.01.
72. Abu Mohammad Eahsan Rasul, Ieyoshi Kobayashi, Tamotsu Kiyoshima, Kou Matsuo, Masamichi Ohishi, Hidetaka Sakai, Expression Pattern of Epstein-Barr Virus Latent Genes in Cell Lines Derived fromOral Squamous Cell Carcinoma., Oral Med. Pathol, 9:95-102, 2004.01.
73. Ogasawara T, Yoshimine Y, Kiyoshima T, Kobayashi I, Matsuo K, Akamine A, Sakai H, In situ expression of RANKL, RANK, osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue., J Periodontal Res., 10.1111/j.1600-0765.2004.00699.x, 39, 1, 42-49, Vol.39, pp.42-9., 2004.01.
74. Kiyoshima T, Yamauchi M, Wong C, Jheon A, Ganss B, Sodek J., An L1 element disrupts human bone sialoprotein promoter: lack of tissue-specific regulation by distalless5 (Dlx5) and runt homeodomain protein2 (Runx2)/core binding factor a1 (Cbfa1) elements., Gene, Vol.299, pp.205-17., 2002.10.
75. Okamura K, Kiyoshima T, Shima K, Kobayashi I, Matsuo K, Ishibashi H, Komatsu S, Rasul AM, Sakai H., Immunohistochemical expression of CA19-9 and CA125 in mucoepidermoid and adenoid cystic carcinomas of the salivary gland., Oral Oncol., 10.1016/S1368-8375(01)00049-5, 38, 3, 244-250, Vol.38, pp.244-50., 2002.04.
76. Wada H, Kobayashi I, Yamaza H, Matsuo K, Kiyoshima T, Akhtar M, Sakai T, Koyano K, Sakai H., In situ expression of heat shock proteins, Hsc73, Hsj2 and Hsp86 in the developing tooth germ of mouse lower first molar., Histochem J., 10.1023/A:1020930228303, 34, 3-4, 105-109, Vol.34, pp.105-9., 2002.03.
77. Yamaza H, Matsuo K, Kobayashi I, Wada H, Kiyoshima T, Akhtar M, Ishibashi Y, Sakai T, Akamine A, Sakai H., Expression of Set-alpha during morphogenesis of mouse lower first molar, Histochem J., 10.1023/A:1014491111628, 33, 8, 437-441, Vol.33,pp.437-41, 2001.08.
78. Kiyoshima T, Shima K, Kobayashi I, Matsuo K, Okamura K, Komatsu S, Rasul AM, Sakai H., Expression of p53 tumor suppressor gene in adenoid cystic and mucoepidermoid carcinomas of the salivary glands, Oral Oncol., 10.1016/S1368-8375(00)00083-X, 37, 3, 315-322, Vol.37,pp.315-22, 2001.04.
79. Shigemura N, Kiyoshima T, Sakai T, Matsuo K, Momoi T, Yamaza H, Kobayashi I, Wada H, Akamine A, Sakai H., Localization of activated caspase-3-positive and apoptotic cells in the developing tooth germ of the mouse lower first molar, Histochem J., 10.1023/A:1017900305661, 33, 5, 253-258, Vol.33,pp.253-8, 2001.03.
80. Yamaza H, Matsuo K, Kiyoshima T, Shigemura N, Kobayashi I, Wada H, Akamime A, Sakai H., Detection of differentially expressed genes in the early developmental stage of the mouse mandible, Int J Dev Biol., 45, 4, 675-680, Vol.45,pp.675-80, 2001.01.
81. Shima K, Kobayashi I, Saito I, Kiyoshima T, Matsuo K, Ozeki S, Ohishi M, Sakai H., Incidence of human papillomavirus 16 and 18 infection and p53 mutation in patients with oral squamous cell carcinoma in Japan, Br J Oral Maxillofac Surg., Vol.38,pp.445-50, 2000.10.
82. Li C.Y., Shirasuna K., Ishibashi H., Nakayama H. and Kiyoshima T., Epithelial-myoepithelial carcinoma arising in pleomorphic adenoma of the palate., Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod., 10.1067/moe.2000.108099, 90, 4, 460-465, 2000.10.
83. Sakai T, Kiyoshima T, Kobayashi I, Moroi R, Ibuki T, Nagadome M, Terada Y, Sakai H., Age-dependent changes in the distribution of BrdU- and TUNEL-positive cells in the murine gingival tissue, J Periodontol., 10.1902/jop.1999.70.9.973, 70, 9, 973-981, Vol.70,pp.973-81, 1999.09.
84. Kobayashi I, Shima K, Saito I, Kiyoshima T, Matsuo K, Ozeki S, Ohishi M, Sakai H., Prevalence of Epstein-Barr virus in oral squamous cell carcinoma, J Pathol., 10.1002/(SICI)1096-9896(199909)189:1<34::AID-PATH391>3.0.CO;2-4, 189, 1, 34-39, Vol.189,pp.34-9, 1999.09.
85. Shigemura N, Kiyoshima T, Kobayashi I, Matsuo K, Yamaza H, Akamine A, Sakai H., The distribution of BrdU- and TUNEL-positive cells during odontogenesis in mouse lower first molars, Histochem J., 10.1023/A:1003796023992, 31, 6, 367-377, Vol.31,pp.367-77, 1999.07.
86. Kobayashi I., Kiyoshima T., Ozeki S., Shima K., Shigemura N., Matsuo K. and Sakai H., Immunohistochemical and ultrastructural study of a papillary cystadenocarcinoma arising from the sublingual gland., J. Oral Pathol. Med., 28, 6, 282-286, 1999.07.
87. Manato JA., Kiyoshima T., Kobayashi I., Shima K., Ohishi M. and Sakai H., The role of macrophages in the absorption process of suture materials: A histological and immunohistochemical study., Acta Histochemica et Cytochemica, 31, 2, 113-120, 1998.03.
88. Kiyoshima T., Kobayashi, I., Matsuo K., Ishibashi Y., Miyoshi A., Akashi Y. and Sakai H., Immunohistochemical localization of laminin, collagen type IV and heparan sulfate proteoglycan in human colorectal adenocarcinoma: correlation with local invasive pattern and lymph node metastasis, Acta Histochemica et Cytochemica, 31, 1, 39-47, Vol.31,pp.39-47, 1998.01.
89. Kazuhiko Okamura, Ieyoshi Kobayashi, Kou Matsuo, Tamotsu Kiyoshima, Kenji Yamamoto, Akira Miyoshi, Hidetaka Sakai, Immunohistochemical localization of cathepsin D, proliferating cell nuclear antigen and epidermal growth factor receptor in human breast carcinoma analysed by computer image analyser: correlation with histological grade and metastatic behaviour, HISTOPATHOLOGY, 31, 6, 540-548, 1997.12.
90. Takayoshi Yamaza, Mizuho A Kido, Tamotsu Kiyoshima, Yukio Nishimura, Masaru Himeno, Teruo Tanaka, A fluid-phase endocytotic capacity and intracellular degradation of a foreign protein (horseradish peroxidase) by lysosomal cysteine proteinases in the rat junctional epithelium, JOURNAL OF PERIODONTAL RESEARCH, DOI: 10.1111/j.1600-0765.1997.tb00575.x, 32, 8, 651-660, 1997.11.
91. Kobayashi I., Matsuo K., Kiyoshima T., Shinohara M. and Sakai H., Salivary Duct Carcinoma with Sebaceous Cell Differentiation arising from Parotid Gland: Histological, Immunohistochemical and Ultrastructural Analyses of a Case., Oral Med. Pathol., 2, 89-93, 1997.03.
92. Okamura K., Kobayashi I., Matsuo K., Kiyoshima T., Yamamoto K., Miyoshi A. and Sakai H., Immunohistochemical localization of cathepsin D, proliferating cell nuclear antigen and epidermal growth factor receptor in human breast carcinoma analyzed by computor image analyzer: correlation with the histological grade and the metastatic activity of carcinoma., Histopathology, 31, 540-548, 1997.03.
93. Teiichi Ibuki, Mizuho A Kido, Tamotsu Kiyoshima, Yoshihiro TERADA, Teruo Tanaka, An ultrastructural study of the relationship between sensory trigeminal nerves and odontoblasts in rat dentin pulp as demonstrated by the anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP), JOURNAL OF DENTAL RESEARCH, 75, 12, 1963-1970, 1996.12.
94. Seiji Shimizu, Mizuho A Kido, Tamotsu Kiyoshima, Teruo Tanaka, Postnatal development of substance P-, calcitonin gene-related peptide- and neuropeptide Y-like immunoreactive nerve fibres in the synovial membrane of the rat temporomandibular joint, ARCHIVES OF ORAL BIOLOGY, 41, 8-9, 749-759, 1996.08.
95. Kenji Murai, Fumitaka Takeshita, Yasunori AYUKAWA, Tamotsu Kiyoshima, Tsuneo Suetsugu, Teruo Tanaka, Light and electron microscopic studies of bone-titanium interface in the tibiae of young and mature rats, JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, 30, 4, 523-533, 1996.04.
96. Kou Matsuo, Ieyoshi Kobayashi, Takayuki TSUKUBA, Tamotsu Kiyoshima, Yukiko Ishibashi, Akira Miyoshi, Kenji Yamamoto, Hidetaka Sakai, Immunohistochemical localization of cathepsins D and E in human gastric cancer: A possible correlation with local invasive and metastatic activities of carcinoma cells, HUMAN PATHOLOGY, 27, 2, 184-190, 1996.02.
97. Teruyoshi Kondo, Mizuho A Kido, Tamotsu Kiyoshima, Takayoshi Yamaza, Teruo Tanaka, AN IMMUNOHISTOCHEMICAL AND MONASTRAL BLUE-VASCULAR LABELING STUDY ON THE INVOLVEMENT OF CAPSAICIN-SENSITIVE SENSORY INNERVATION OF THE JUNCTIONAL EPITHELIUM IN NEUROGENIC PLASMA EXTRAVASATION IN THE RAT GINGIVA, ARCHIVES OF ORAL BIOLOGY, DOI: 10.1016/0003-9969(95)00060-3, 40, 10, 931-940, 1995.10.
98. Ryoji Moroi, Takayoshi Yamaza, Yasunori AYUKAWA, Tamotsu Kiyoshima, Yasuyoshi Ohsaki, Yukio Nishimura, Yoshihiro TERADA, Masaru Himeno, Teruo Tanaka, Changes in the immunocytochemical localization of cathepsin L and type I collagen in rat osteoclasts treated with E-64, ACTA HISTOCHEMICA ET CYTOCHEMICA, 28, 6, 523-531, 1995.12.
99. Mizuho A Kido, S Shimizu, Tamotsu Kiyoshima, Yoshihiro TERADA, Teruo Tanaka, Neuropeptide Y-, tyrosine hydroxylase-, vasoactive intestinal polypeptide-immunoreactive nerves in TMJ., JOURNAL OF DENTAL RESEARCH, 75, 749, 1996.01.
100. Mizuho A Kido, Tamotsu Kiyoshima, teiichi ibuki, S Shimizu, Teruyoshi Kondo, Yoshihiro TERADA, Teruo Tanaka, A TOPOGRAPHICAL AND ULTRASTRUCTURAL-STUDY OF SENSORY TRIGEMINAL NERVE-ENDINGS IN THE RAT TEMPOROMANDIBULAR-JOINT AS DEMONSTRATED BY ANTEROGRADE TRANSPORT OF WHEAT-GERM AGGLUTININ-HORSERADISH PEROXIDASE (WGA-HRP), JOURNAL OF DENTAL RESEARCH, 74, 7, 1353-1359, 1995.01.
101. Tamotsu Kiyoshima, Mizuho A Kido, Yukio Nishimura, Masaru Himeno, Takayuki TSUKUBA, Hideo Tashiro, Kenji Yamamoto, Teruo Tanaka, IMMUNOCYTOCHEMICAL LOCALIZATION OF CATHEPSIN-L IN THE SYNOVIAL LINING CELLS OF THE RAT TEMPOROMANDIBULAR-JOINT, ARCHIVES OF ORAL BIOLOGY, 39, 12, 1049-1056, 1994.12.
102. Kou Matsuo, Yukiko Ishibashi, Ieyoshi Kobayashi, Satoru Ozeki, Masamichi OHISHI, T Tange, J Hirata, Tamotsu Kiyoshima, Hidetaka Sakai, NEW HUMAN ORAL SQUAMOUS CARCINOMA CELL-LINE AND ITS TUMORIGENIC SUBLINE PRODUCING GRANULOCYTE-COLONY-STIMULATING FACTOR, JAPANESE JOURNAL OF CANCER RESEARCH, 85, 12, 1257-1262, 1994.12.
103. Tamotsu Kiyoshima, Mizuho A Kido, Takayuki TSUKUBA, Hidetaka Sakai, Kenji Yamamoto, Teruo Tanaka, LOCALIZATION OF CATHEPSIN-B AND CATHEPSIN-D IN THE SYNOVIAL LINING CELLS OF THE NORMAL RAT TEMPOROMANDIBULAR-JOINT BY IMMUNO-LIGHT AND IMMUNO-ELECTRON MICROSCOPY, Acta Histochem Cytochem., 27, 5, 441-450, 1994.10.
104. Emi Nagata, Teruyoshi Kondo, Tamotsu Kiyoshima, Minoru Nakata, Teruo Tanaka, Immunohistochemical evidence for the presence of nerve fibres with substance P- or calcitonin generated peptide-like immunoreactivity in the proliferating epithelium in the developing teeth of rats., Arch Oral Biol., 39, 3, 197-203, 1994.03.
105. Tetsuya Goto, Tamotsu Kiyoshima, Ryoji Moroi, Takayuki TSUKUBA, Yukio Nishimura, Masaru Himeno, Kenji Yamamoto, Teruo Tanaka, Localization of cathepsins B, D, and L in the rat osteoclast by immuno-light and -electron microscopy., Histochem., 101, 1, 33-40, 1994.01.
106. Tamotsu Kiyoshima, Takayuki TSUKUBA, Mizuho A Kido, Hideo Tashiro, Kenji Yamamoto, Teruo Tanaka, Immunohistochemical localization of cathepsins B and D in the synovial lining cells of the normal rat temporomandibular joint., Arch Oral Biol., 38, 4, 357-359, 1993.04.
107. Mizuho A Kido, Tamotsu Kiyoshima, Teruyoshi Kondo, Norio Ayasaka, Ryoji Moroi, Yoshihiro TERADA, Teruo Tanaka, DISTRIBUTION OF SUBSTANCE-P AND CALCITONIN GENE-RELATED PEPTIDE-LIKE IMMUNOREACTIVE NERVE-FIBERS IN THE RAT TEMPOROMANDIBULAR-JOINT, JOURNAL OF DENTAL RESEARCH, 72, 3, 592-598, 1993.03.
108. Tetsuya Goto, Takayuki TSUKUBA, Tamotsu Kiyoshima, Yukio Nishimura, Keitaro Kato, Kenji Yamamoto, Teruo Tanaka, Immunohistochemical localization of cathepsins B, D and L in the rat osteoclast., Histochemistry, 99, 5, 411-414, 1993.05.