TGF-β_(2)对小梁网细胞上皮-间质转化的影响及其分子机制研究

TGF-β_(2)promotes epithelial mesenchymal transformation in human trabecular meshwork cells and its mechanism

目的探讨转化生长因子β_(2)(TGF-β_(2))高表达对小梁网细胞上皮-间质转化(EMT)的影响及作用机制。方法取小梁网细胞系分为对照组和TGF-β_(2)诱导组,分别用DMEM培养液和含不同浓度TGF-β_(2)的DMEM培养液培养,RT-qPCR和Westernblot检测EMT相关指标(α-SMA、VIM、snail)以及TGF-β/Smad信号通路相关因子Smad3、p-Smad3、Smad7的mRNA和蛋白表达水平;划痕实验检测TGF-β_(2)诱导后HTM细胞迁移能力。结果经TGF-β_(2)诱导后,上皮细胞标志物E-cadherinmRNA和蛋白表达水平明显降低,间质细胞标志物(α-SMA、VIM、snail)mRNA和蛋白表达水平明显升高(P<0.01)。经TGF-β_(2)诱导后,TGF-β/Smad信号通路相关因子p-Smad3蛋白表达水平显著升高,Smad7的mRNA显著升高,但Smad7蛋白表达水平无显著变化。划痕实验检测结果显示,TGF-β_(2)诱导后HTM细胞迁移能力显著增强。结论TGF-β_(2)可诱导HTM细胞系发生EMT改变,增强细胞迁移能力,其机制可能与激活TGF-β/Smad信号通路有关。

Objective To investigate the effect of overexpression of TGF-β_(2)on epithelial-mesenchymal transformation in the human trabecular meshwork cells and its molecular mechanism.Methods The human trabecular meshwork cell line was used as the research object.The cells cultured with DMEM medium were served as the control group and the cells cultured with DMEM medium containing 10 ng/mL TGF-β_(2)were served as the TGF-β_(2)group.RT-qPCR and Western blot were used to detect the mRNA and protein expression levels of EMT related indicators(α-SMA,VIM,snail)and TGF-β/Smad signaling pathway related indicators Smad3、p-Smad3、Smad7.Scratch assay was used to detect cell migration ability of HTM cells induced by TGF-β_(2).Results After TGF-β_(2)induction,the epithelial cell marker E-cadherin mRNA and protein expression levels in experimental group significantly decreased,while the mesenchymal cell markers(α-SMA,VIM,snail)mRNA and protein levels significantly increased,and the differences were statistically significant(P<0.01).After TGF-β_(2)induction,the related indicators of TGF-β/Smad signaling pathway,p-Smad3 protein expression level was significantly increased,and Smad7 mRNA was significantly increased,while Smad7 protein expression level did not change significantly.The migration ability of HTM cells was significantly enhanced after TGF-β_(2)induction(P<0.05).Conclusion Overexpression of TGF-β_(2)could induce EMT of human trabecular meshwork cells,and enhance cell migration ability,the mechanism may be related to activation of TGF-β/Smad signaling pathway.