title: Arecoline-induced myofibroblast transdifferentiation from human buccal mucosal fibroblasts is mediated by ZEB1.
abstract: Oral submucous fibrosis (OSF) is considered as a pre-cancerous condition of the oral mucosa and is highly associated with habitual areca quid chewing. Arecoline is the major alkaloid in areca quid and is thought to be involved in the pathogenesis of OSF. Our previous studies have demonstrated that arecoline could induce epithelial-mesenchymal transition (EMT)-related factors in primary human buccal mucosal fibroblasts (BMFs). Therefore, we investigated the expression of zinc finger E-box binding homeobox 1 (ZEB1), which is a well-known transcriptional factor in EMT, in OSF tissues and its role in arecoline-induced myofibroblast transdifferentiation from BMFs. The expression of ZEB1, as well as the myofibroblast marker α-smooth muscle actin (α-SMA), was significantly increased in OSF tissues, respectively. With immunofluorescence analysis, arecoline induced the formation of α-SMA-positive stress fibres in BMFs expressing nuclear ZEB1. Arecoline also induced collagen contraction of BMFs in vitro. By chromatin immunoprecipitation, the binding of ZEB1 to the α-SMA promoter in BMFs was increased by arecoline. The promoter activity of α-SMA in BMFs was also induced by arecoline, while knockdown of ZEB1 abolished arecoline-induced α-SMA promoter activity and collagen contraction of BMFs. Long-term exposure of BMFs to arecoline induced the expression of fibrogenic genes and ZEB1. Silencing of ZEB1 in fibrotic BMFs from an OSF patient also suppressed the expression of α-SMA and myofibroblast activity. Inhibition of insulin-like growth factor receptor-1 could suppress arecoline-induced ZEB1 activation in BMFs. Our data suggest that ZEB1 may participate in the pathogenesis of areca quid-associated OSF by activating the α-SMA promoter and inducing myofibroblast transdifferentiation from BMFs.
authors.affiliation: School of Dentistry, Chung Shan Medical University, Taichung, Taiwan; Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan.
authors.forename: Yu-Chao
authors.initials: YC
authors.lastname: Chang
authors.forename: Chung-Hung
authors.initials: CH
authors.lastname: Tsai
authors.forename: You-Liang
authors.initials: YL
authors.lastname: Lai
authors.forename: Cheng-Chia
authors.initials: CC
authors.lastname: Yu
authors.forename: Wan-Yu
authors.initials: WY
authors.lastname: Chi
authors.forename: Jung Jung
authors.initials: JJ
authors.lastname: Li
authors.forename: Wen-Wei
authors.initials: WW
authors.lastname: Chang
mesh_terms: D000199:Actins / Q000096:biosynthesis / Q000235:genetics; D001114:Areca / Q000737:chemistry; D001115:Arecoline / Q000008:administration & dosage*; D054338:Cell Transdifferentiation / Q000187:drug effects*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D005347:Fibroblasts / Q000187:drug effects; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008409:Mastication; D009061:Mouth Mucosa / Q000187:drug effects / Q000473:pathology; D058628:Myofibroblasts / Q000187:drug effects; D009914:Oral Submucous Fibrosis / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology*; D061251:Primary Cell Culture; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D018398:Homeodomain Proteins; D014157:Transcription Factors; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D001115:Arecoline
keywords: ZEB1; arecoline; buccal mucosal fibroblasts; myofibroblasts; oral submucous fibrosis; α-SMA