Negative feedback by SNAI2 regulates TGFβ1‐induced amelotin gene transcription in epithelial–mesenchymal transition

• Published in: Journal of cellular physiology 2019-07, Vol.234 (7), p.11474-11489
• Main Authors: Nakayama, Yohei, Tsuruya, Yuto, Noda, Keisuke, Yamazaki‐Takai, Mizuho, Iwai, Yasunobu, Ganss, Bernhard, Ogata, Yorimasa
• Format: Article
• Language: English
• Subjects: amelotin; Animals; Apoptosis; Cell Line; Cytokines; Dental Enamel Proteins - genetics; Dental Enamel Proteins - metabolism; Down-Regulation; Epithelial cells; Epithelial Cells - metabolism; Epithelial-Mesenchymal Transition - physiology; epithelial–mesenchymal transition; Epithelium; Feedback; Fibrosis; Gene expression; Gene Expression Regulation - drug effects; Gingiva; Gingiva - cytology; Growth factors; Homeostasis; Inflammation; junctional epithelium; Mesenchyme; Mice; Negative feedback; Periodontics; Ribonucleic acid; RNA; Signal transduction; Signaling; siRNA; Smad3 protein; SNAI2; Snail Family Transcription Factors - genetics; Snail Family Transcription Factors - metabolism; Snail protein; Transcription; Transfection; Transforming Growth Factor beta1 - pharmacology; Wound healing
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.27804

Junctional epithelium (JE) demonstrates biological responses with the rapid turnover of gingival epithelial cells. The state occurs in inflammation of gingiva and wound healing after periodontal therapy. To understand the underlying mechanisms and to maintain homeostasis of JE, it is important to investigate roles of JE‐specific genes. Amelotin (AMTN) is localized at JE and regulated by inflammatory cytokines and apoptotic factors that represent a critical role of AMTN in stabilizing the dentogingival attachment, which is an entrance of oral bacteria. In this study, we demonstrated that the AMTN gene expression was regulated by SNAI2 and transforming growth factor β1 (TGFβ1)‐induced epithelial–mesenchymal transition (EMT) that occurs in wound healing and fibrosis during chronic inflammation. SNAI2 downregulated AMTN gene expression via SNAI2 bindings to E‐boxes (E2 and E4) in the mouse AMTN gene promoter in EMT of gingival epithelial cells. Meanwhile, TGFβ1‐induced AMTN gene expression was attenuated by SNAI2 and TGFβ1‐induced SNAI2, without inhibition of the TGFβ1‐Smad3 signaling pathway. Moreover, SNAI2 small interfering RNA (siRNA) rescued SNAI2‐induced downregulation of AMTN gene expression, and TGFβ1‐induced AMTN gene expression was potentiated by SNAI2 siRNA. Taken together, these data demonstrated that AMTN gene expression in the promotion of EMT was downregulated by SNAI2. The inhibitory effect of AMTN gene expression was an independent feedback on the TGFβ1‐Smad3 signaling pathway, suggesting that the mechanism can be engaged in maintaining homeostasis of gingival epithelial cells at JE and the wound healing phase. Transforming growth factor β1 (TGFβ1) upregulated amelotin (AMTN gene) expression in gingival epithelial cells. TGFβ1‐induced AMTN gene expression was subsequently downregulated by TGFβ1‐induced SNAI2 at epithelial–mesenchymal transition (EMT), mediated by SNAI2 bindings to E‐boxes in AMTN gene promoter. These results suggested that AMTN plays a role in EMT to maintain homeostasis of junctional epithelium.