miR-200a在TGF-β诱导口腔黏膜修复中的功能研究

Functional Studies of miR-200a in TGF-β Induced Oral Mucosal Repair

目的探索miR-200a在TGF-β诱导口腔黏膜修复过程中表达水平的变化和功能。方法分离培养原代人牙龈成纤维细胞(Human Gingival Fibroblast,HGF),加入不同浓度TGF-β,实时荧光定量PCR检测miR-200表达水平的变化。通过转染miR-200a模拟物增加miR-200a表达水平,利用Transwell实验检测细胞迁移能力,利用CCK-8实验检测细胞增殖活性,利用蛋白质免疫印迹实验检测细胞Ⅰ型胶原和Ⅲ型胶原表达水平。结果TGF-β处理会引起HGF细胞miR-200a表达水平下降,且随着TGF-β作用浓度增加,miR-200a表达下降比例增加;与对照组相比,转染miR-200a模拟物的实验组miR-200a表达水平上升,细胞增殖活性下降,迁移到Transwell下层小室的细胞数目降低,Ⅰ型和Ⅲ型胶原蛋白表达水平下降。结论miR-200a在TGF-β诱导口腔黏膜修复过程中可以抑制HGF的增殖活性、迁移能力和Ⅰ型、Ⅲ型胶原蛋白的表达,进而影响口腔黏膜的修复和重建。

Objective To explore the expression and function of miR-200a in TGF-βinduced oral mucosal repair.Methods Human primary gingival fibroblasts(HGF)were isolated and cultured,then TGF-βwith different concentration was added to detect the expression of miR-200a by Real-time PCR assay.The miR-200a expression was up-regulated by transfection with miR-200a mimics.The cell proliferation was measured by CCK-8 assay;The cell migration rate was measured by Transwell assay;The expressions of CollagenⅠand CollagenⅢwere detected by western blot.Results The miR-200a expression levels in HGF cells treated with TGF-βwere decreased,and the decreased rates were related with TGF-βconcentration;Compared with control group,the experimental group transfected with miR-200a mimics showed increased miR-200a expression,decreased cell proliferation activity,decreased cell number migrating to the lower compartment of Transwell,and decreased collagen typeⅠandⅢexpression levels.Conclusion miR-200a can inhibit the proliferation activity,migration ability and expression of typeⅠandⅢcollagen of HGF during TGF-βinduced oral mucosal repair,thus affecting the repair and reconstruction of oral mucosa.