TGF-β2 promotes epithelial mesenchymal transformation in human trabecular meshwork cells and its mechanism
Objective To investigate the effect of overexpression of TGF-β2 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-β2 were served as the TGF-β2 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-β2.Results After TGF-β2 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-β2 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-β2 induction(P＜0.05).Conclusion Overexpression of TGF-β2 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.
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