β-CATENIN REGULATES Wnt5a AND PARTICIPATES IN EPITHELIAL MESENCHYMAL TRANSDIFFERENTIATION OF HUMAN LENS EPITHELIAL CELLS
Objective To observe the role of β-catenin in the regulation of Wnt5a expression by transform growth factor-β2(TGF-β2)and its effect on TGF-β2-induced epithelial mesenchymal transition(EMT).Methods Take human lens epithelial cells(SRA01/04 cells)which were cultured in vitro as research object,added exogenous 10 ng/mL TGF-β2 to induce cellular EMT.48 hours prior to the addition of TGF-β2,liposome-mediated transfection technology was employed as an intervention treatment.This process involved the transient transfection of β-catenin siRNA or the S33Y-β-catenin cDNA expression vector for descending or over expression of β-catenin.Western Blot was applied to detect the expression of β-catenin,Wnt5a,E-cad-herin and α-SMA proteins in each group.Results After the addition of TGF-β2,the expression of Wnt5a protein and β-catenin protein in SRA01/04 cells was significantly up-regulated;the relative expression of E-cadherin protein decreased,and the rel-ative expression of α-SMA protein increased.After transfection with β-catenin-siRNA,the expression of β-catenin protein was significantly down-regulated,while there was no significant change in the expression of Wnt5a protein;After stimulation with β-catenin-siRNA+TGF-β 2,the expression of E-cadherin protein was up-regulated,whereas the expression of β-catenin,Wnt5a and a-SMA proteins were down-regulated.After transfection of S33Y-β-catenin cDNA,the expression of β-catenin protein was significantly elevated,whereas there was no significant change in the expression of Wnt5a protein.α-SMA protein expression was significantly up-regulated and E-cadherin protein expression was down-regulated after stimulation with S33Y-β-catenin+TGF-β2.Conclusions TGF-β2 induced overexpression of Wnt5a and β-catenin in normal lens epithe-lial cells,and induced EMT;overexpression or degradation of β-catenin can promote or decrease the expression of TGF-β2-in-duced Wnt5a,and participate in TGF-β2-induced EMT.
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