Kyushu University Academic Staff Educational and Research Activities Database
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Shinsuke Fujii Last modified date:2024.06.03

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

Graduate School
Undergraduate School

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 Reseacher Profiling Tool Kyushu University Pure
Academic Degree
Country of degree conferring institution (Overseas)
Field of Specialization
Morphological Oral Biology, Oral pathology
Total Priod of education and research career in the foreign country
Research Interests
  • The expression of Arl4c and its function in tumorigenesis.
    keyword : Arl4c, Tumorigenesis
  • The effect of Wnt signaling on salivary gland development.
    keyword : Wnt signaling, salivary gland
  • The effects of TGF-β1 on the proliferation and differentiation of human periodontal ligament cells
    keyword : TGF-β1、periodontal ligament cell
  • Establishment of human periodontal ligament stem/progenitor cell lines and its characterization
    keyword : stem/progenitor cell, periodontal ligament cell
  • Immortalization of human periodontal ligament cells
    keyword : Immortalization. Periodontal ligament
    2003.04~2006.03I established human periodontal ligament progenitor cell lines for the study of regeneration of human periodontal ligament. I am trying to clarify the regeneration mechanism of human periodontal ligament using human periodontal ligament progenitor cell lines..
Academic Activities
1. Ryoko Nagano, Yusuke Nakako, Shinsuke Fujii, Shintaro Kawano, Hidefumi Maeda, Tamotsu Kiyoshima, The IL-1β-p65 axis stimulates quiescent odontogenic epithelial cell rests via TGF-β signalling to promote cell proliferation of the lining epithelia in radicular cysts: A laboratory investigation., International endodontic journal, 10.1111/iej.14016, 57, 3, 344-354, 2024.03, AIM: Cyst formation of the jaws is frequently accompanied by the proliferation of odontogenic epithelial cells located in the periodontal ligament (PDL), which consists of heterozygous cells and includes the most fibroblasts. The lining epithelium of radicular cyst, an odontogenic cyst of inflammatory origin, is derived from the proliferation of the remnants of the Hertwig epithelial root sheath (odontogenic epithelial cell rests of Malassez; ERMs) in the PDL. ERMs are maintained at a lower proliferative state under physiological conditions, but the regulatory mechanisms underlying the inflammation-dependent enhanced-proliferative capabilities of ERMs are not fully understood. The aim of this study was to evaluate the effects of cytokine pathway association between TGF-β signalling and IL-1β signalling on the regulation of odontogenic epithelial cell proliferation using radicular cyst pathological specimens and odontogenic epithelial cell lines. METHODOLOGY: Immunofluorescence analyses were performed to clarify the expression levels of Smad2/3 and Ki-67 in ERMs of 8-week-old mouse molar specimens. In radicular cyst (n = 52) and dentigerous cysts (n = 6) specimens from human patients, the expression of p65 (a main subunit of NF-κB), Smad2/3 and Ki-67 were investigated using immunohistochemical analyses. Odontogenic epithelial cells and PDL fibroblastic cells were co-cultured with or without an inhibitor or siRNAs. Odontogenic epithelial cells were cultured with or without TGF-β1 and IL-1β. The proliferative capabilities and Smad2 phosphorylation levels of odontogenic epithelial cells were examined. RESULTS: Immunohistochemically, Smad2/3-positivity was increased, and p65-positivity and Ki-67-positivity were decreased both in ERMs and in the epithelial cells in dentigerous cysts, a non-inflammatory developmental cyst. In contrast, p65-positive cells, along with the expression of Ki-67, were increased and Smad2/3-positive cells were decreased in the lining epithelia of radicular cysts. Co-culture experiments with odontogenic epithelial cells and PDL fibroblastic cells revealed that PDL cells-derived TGF-β1/2 and their downstream signalling suppressed odontogenic epithelial cell proliferation. Moreover, TGF-β1 stimulation induced Smad2 phosphorylation and suppressed odontogenic epithelial cell proliferation, while IL-1β stimulation reversed these phenotypes through p65 transactivation. CONCLUSIONS: These results suggest that IL-1β-p65 signalling promotes odontogenic epithelial cell proliferation through suppressing TGF-β-Smad2 signalling, which would be involved in the pathogenesis of radicular cysts..
2. Shinsuke Fujii, Kana Hasegawa, Takashi Maehara, Kari J Kurppa, Kristiina Heikinheimo, Kristy A. Warner, Satoshi Maruyama, Yudai Tajiri, Jacques E. Nör, Jun-ichi Tanuma, Shintaro Kawano, Tamotsu Kiyoshima, Wnt/β-catenin-C-kit axis may play a role in adenoid cystic carcinoma prognostication, Pathology - Research and Practice, 10.1016/j.prp.2024.155148, 155148-155148, 2024.01.
3. Thinh Thi Kim Truong, Shinsuke Fujii, Ryoko Nagano, Kana Hasegawa, Megumi Kokura, Yuta Chiba, Keigo Yoshizaki, Satoshi Fukumoto, Tamotsu Kiyoshima, Arl4c is involved in tooth germ development through osteoblastic/ameloblastic differentiation., Biochemical and biophysical research communications, 10.1016/j.bbrc.2023.09.014, 679, 167-174, 2023.09, Murine tooth germ development proceeds in continuous sequential steps with reciprocal interactions between the odontogenic epithelium and the adjacent mesenchyme, and several growth factor signaling pathways and their activation are required for tooth germ development. The expression of ADP-ribosylation factor (Arf)-like 4c (Arl4c) has been shown to induce cell proliferation, and is thereby involved in epithelial morphogenesis and tumorigenesis. In contrast, the other functions of Arl4c (in addition to cellular growth) are largely unknown. Although we recently demonstrated the involvement of the upregulated expression of Arl4c in the proliferation of ameloblastomas, which have the same origin as odontogenic epithelium, its effect on tooth germ development remains unclear. In the present study, single-cell RNA sequencing (scRNA-seq) analysis revealed that the expression of Arl4c, among 17 members of the Arf-family, was specifically detected in odontogenic epithelial cells, such as those of the stratum intermedium, stellate reticulum and outer enamel epithelium, of postnatal day 1 (P1) mouse molars. scRNA-seq analysis also demonstrated the higher expression of Arl4c in non-ameloblast and inner enamel epithelium, which include immature cells, of P7 mouse incisors. In the mouse tooth germ rudiment culture, treatment with SecinH3 (an inhibitor of the ARNO/Arf6 pathway) reduced the size, width and cusp height of the tooth germ and the thickness of the eosinophilic layer, which would involve the synthesis of dentin and enamel matrix organization. In addition, loss-of-function experiments using siRNAs and shRNA revealed that the expression of Arl4c was involved in cell proliferation and osteoblastic cytodifferentiation in odontogenic epithelial cells. Finally, RNA-seq analysis with a gene set enrichment analysis (GSEA) and Gene Ontology (GO) analysis showed that osteoblastic differentiation-related gene sets and/or GO terms were downregulated in shArl4c-expressing odontogenic epithelial cells. These results suggest that the Arl4c-ARNO/Arf6 pathway axis contributes to tooth germ development through osteoblastic/ameloblastic differentiation..
4. Shinsuke Fujii, Tamotsu Kiyoshima, The role of Wnt, ARL4C, and Sema3A in developmental process and disease pathogenesis., Pathology international, 10.1111/pin.13325, 73, 6, 217-233, 2023.06, Various types of tumors, including malignant and benign ones, occur in the oral cavity. These arise from the mucosal epithelium, odontogenic epithelium, and salivary gland. To date, few major driver events in oral tumors have been identified. Accordingly, molecular targets in anti-tumor therapy for oral tumors are lacking. We focused on elucidating the function of aberrantly activated signal transduction related to oral tumor formation, especially in oral squamous cell carcinoma, ameloblastoma, and adenoid cystic carcinoma, which are raised as common oral tumors. Wnt/β-catenin-dependent pathway is involved in the developmental process, organ homeostasis and disease pathogenesis through regulating various cellular functions by enhancing transcriptional activity. Recently, we identified ADP-ribosylation factor (ARF)-like 4c (ARL4C) and Semaphorin 3A (Sema3A), the expression of which is regulated by Wnt/β-catenin-dependent pathway, and characterized their functions in the developmental process and tumor formation. This review highlights the recent advances in understanding the roles of Wnt/β-catenin-dependent pathway, ARL4C and Sema3A, as determined by pathological and experimental studies..
5. Dania Zuhier Ragheb Alkhatib, Thinh Thi Kim Truong, Shinsuke Fujii, Kana Hasegawa, Ryoko Nagano, Yudai Tajiri, Tamotsu Kiyoshima, Stepwise activation of p63 and the MEK/ERK pathway induces the expression of ARL4C to promote oral squamous cell carcinoma cell proliferation., Pathology, research and practice, 10.1016/j.prp.2023.154493, 246, 154493-154493, 2023.06, Carcinogenesis is a multistep process wherein cells accumulate multiple genetic alterations and progress to a more malignant phenotype. It has been proposed that sequential accumulation of gene abnormalities in specific genes drives the transition from non-tumorous epithelia through a preneoplastic lesion/benign tumor to cancer. Histologically, oral squamous cell carcinoma (OSCC) progresses in multiple ordered steps that begin with mucosal epithelial cell hyperplasia, which is followed by dysplasia, carcinoma in situ and invasive carcinoma. It is therefore hypothesized that genetic alteration-mediated multistep carcinogenesis would be involved in the development of OSCC; however, the detailed molecular mechanisms are unknown. We clarified the comprehensive gene expression patterns and carried out an enrichment analysis using DNA microarray data from a pathological specimen of OSCC (including a non-tumor region, carcinoma in situ lesion and invasive carcinoma lesion). The expression of numerous genes and signal activation were altered in the development of OSCC. Among these, the p63 expression was increased and the MEK/ERK-MAPK pathway was activated in carcinoma in situ lesion and in invasive carcinoma lesion. Immunohistochemical analyses revealed that p63 was initially upregulated in carcinoma in situ and ERK was sequentially activated in invasive carcinoma lesions in OSCC specimens. ADP-ribosylation factor (ARF)-like 4c (ARL4C), the expression of which is reportedly induced by p63 and/or the MEK/ERK-MAPK pathway in OSCC cells, has been shown to promote tumorigenesis. Immunohistochemically, in OSCC specimens, ARL4C was more frequently detected in tumor lesions, especially in invasive carcinoma lesions, than in carcinoma in situ lesions. Additionally, ARL4C and phosphorylated ERK were frequently merged in invasive carcinoma lesions. Loss-of-function experiments using inhibitors and siRNAs revealed that p63 and MEK/ERK-MAPK cooperatively induce the expression of ARL4C and cell growth in OSCC cells. These results suggest that the stepwise activation of p63 and MEK/ERK-MAPK contributes to OSCC tumor cell growth through regulation of ARL4C expression..
6. Nagano, Ryoko; Fujii, Shinsuke; Hasegawa, Kana; Maeda, Hidefumi; Kiyoshima, Tamotsu, Wnt signaling promotes tooth germ development through YAP1-TGF- βsignaling, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 10.1016/j.bbrc.2022.09.012, 630, 64-70, 2022.11.
7. Yurie Mikami, Shinsuke Fujii, Kenichi Kouhashi, Yuichi Yamada, Masafumi Moriyama, Shintarou 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.
8. Mikami Yurie, Fujii Shinsuke, 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, 143, 8, 1381-1393, 2017.03.
9. Fujii S, Shinjo K, Matsumoto S, Harada T, Nojima S, Sato S, Usami Y, Toyosawa S, Morii E, Kondo Y, Kikuchi A, Epigenetic upregulation of ARL4C, due to DNA hypomethylation in the 3’-untranslated region, promotes tumorigenesis of lung squamous cell carcinoma, Oncotarget, doi: 10.18632/oncotarget.13147., 6, 49, 81571-81587, 2016.10.
10. Fujii S, Maeda H, Wada N, Tomokiyo A, Saito M, Akamine A, Investigating a clonal human periodontal ligament progenitor/stem cell line in vitro and in vivo., J Cell Physiol, 215(3):743-749, 2008.03.
11. Shinsuke Fujii., Hidefumi Maeda., Naohisa Wada., Yoshio Kano., Akifumi Akamine. , Establishing and characterizing human periodontal ligament fibroblasts immortalized by SV40T-antigen and hTERT gene transfer., Cell and tissue research, 324巻 117-125頁, 2006.01.
1. Shinsuke Fujii, Abnormality of Signal Network in Oral Tumor, IAOP 2023, 21st International Congress of Oral Pathology and Medicine, 2023.08.
2. Tatsufumi Fujimoto, Shinsuke Fujii, Tamotsu Kiyoshima, Sema3A-AKT Axis In Salivary Gland And Adenoid Cystic Carcinoma Developments, 第68回JADR学術大会, 2020.11.
Membership in Academic Society
Other Educational Activities
  • 2016.05.
  • 2009.11.
  • 2008.11.
  • 2007.11.