School of Dentistry, Chung Shan Medical University, Taichung, Taiwan; Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan.
School of Dentistry, Chung Shan Medical University, Taichung, Taiwan; Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan; Institute of Oral Sciences, Chung Shan Medical University, Taichung, Taiwan.
J Formos Med Assoc. 2018 Sep;117(9):791-797. doi: 10.1016/j.jfma.2018.05.016. Epub 2018 Jun 27.
BACKGROUND/PURPOSE: MicroRNA-200c (miR-200c) recently emerged as an important regulator of tumorigenesis and cancer metastasis, however, its role in regulating oral submucous fibrosis (OSF) remains unknown. In this study, we investigated the functional role of miR-200c in myofibroblastic differentiation activity and identified its potential target.
qRT-PCR was applied to assess the expression of miR-200c in OSF tissues and fibrotic buccal mucosal fibroblasts (fBMFs). Arecoline, a major areca nut alkaloid, was utilized to explore whether the expression of miR-200c would alter following stimulation. Collagen gel contraction, migration and invasion capabilities were examined in arecoline-stimulated BMFs as wells as in fBMFs. Luciferase reporter assay was conducted to show the relationship between miR-200c and ZEB1.
Our results showed that the expression of miR-200c was downregulated in OSF specimen and fBMFs. Arecoline treatment dose-dependently reduced the relative expression of miR-200c in normal BMFs. Overexpression of miR-200c impeded the arecoline-induced collagen gel contraction, migration, invasion and wound healing capacities. Moreover, ectopic expression of miR-200c in fBMFs successfully reduced the increased collagen gel contractility and invasion abilities. Our results demonstrated that ZEB1 was a direct target of miR-200c, and overexpression of miR-200c inhibited the expression of ZEB1 and α-SMA.
These findings suggest that downregulation of miR-200c in OSF may be involved in the pathogenesis of areca nut-associated OSF through regulation of ZEB1.
背景/目的：微小 RNA-200c（miR-200c）最近成为肿瘤发生和癌症转移的重要调节剂，然而，其在调节口腔黏膜下纤维性变（OSF）中的作用尚不清楚。在这项研究中，我们研究了 miR-200c 在肌成纤维细胞分化活性中的功能作用，并鉴定了其潜在的靶标。
应用 qRT-PCR 评估 miR-200c 在 OSF 组织和纤维化颊黏膜成纤维细胞（fBMFs）中的表达。利用槟榔碱，一种主要的槟榔生物碱，来探讨 miR-200c 的表达是否会在刺激后发生改变。在槟榔碱刺激的 BMFs 以及 fBMFs 中检测胶原凝胶收缩、迁移和侵袭能力。通过荧光素酶报告基因实验显示 miR-200c 与 ZEB1 之间的关系。
我们的结果表明，miR-200c 在 OSF 标本和 fBMFs 中的表达下调。槟榔碱处理剂量依赖性地降低了正常 BMFs 中 miR-200c 的相对表达。miR-200c 的过表达阻碍了槟榔碱诱导的胶原凝胶收缩、迁移、侵袭和伤口愈合能力。此外，在 fBMFs 中外源性表达 miR-200c 成功降低了增加的胶原凝胶收缩性和侵袭能力。我们的结果表明，ZEB1 是 miR-200c 的直接靶标，过表达 miR-200c 抑制了 ZEB1 和 α-SMA 的表达。
这些发现表明，OSF 中 miR-200c 的下调可能通过调节 ZEB1 参与槟榔相关 OSF 的发病机制。
