期刊
ARCHIVES OF ORAL BIOLOGY
卷 71, 期 -, 页码 46-53出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.archoralbio.2016.06.025
关键词
Gingival epithelial cells; Cell adhesion; Smad2/3; Integrin
资金
- JSPS [22890119, 24792327, 15K20620]
- Grants-in-Aid for Scientific Research [25253104, 22890119, 15K11404, 24792327, 15K20620] Funding Source: KAKEN
Objective: Gingival epithelial cells play an important role in preventing the initiation of periodontitis, by their hemidesmosomal adhesion to the tooth root surface. Adhesion requires integrin-extracellular matrix (ECM) interactions that are intricately regulated by transforming growth factor-beta (TGF-beta) signaling. However, the mechanisms underlying the interplay between adhesion molecules and TGF-beta, especially the respective roles of Smad2 and Smad3, remain elusive. In this study, we examined the effects of Smad overexpression on gingival epithelial cell adhesion and expression profiles of integrin and ECM-related genes. Methods: Human gingival epithelial cells immortalized by the SV40 T-antigen were transfected with Smad2- and Smad3-overexpression vectors. A cell adhesion assay involving fluorescence detection of attached cells was performed using the ArrayScan imaging system. Real-time PCR was performed to examine the kinetics of integrin and ECM gene expression. In vitro and in vivo localization of adhesion molecules was examined by immunofluorescence analysis. Results: By using SB431542, a specific inhibitor of the TGF-beta type I receptor, Smad2/3 signaling was confirmed to be dominant in TGF-beta 1-induced cell adhesion. The Smad2-transfectant demonstrated higher potency for cell adhesion and integrin expression (alpha 2, alpha 5, beta 4, and beta 6) than the Smad3-transfectant, whereas little or no change in ECM expression was observed in either transfectant. Moreover, the gingival epithelium of transgenic mice that overexpressed Smad2 driven by the keratin 14 promoter showed increased integrin alpha 2 expression. Conclusion: These findings indicate the crucial role of Smad2 in increased adhesion of gingival epithelial cells via upregulation of integrin alpha 2. (C) 2016 Elsevier Ltd. All rights reserved.
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