LncRNA-AV310809 Promotes TGF-β1-induced Epithelial Mesenchymal Transition(EMT) Of Peritoneal Mesothelial Cells Via Wnt2/β-catenin Signaling Pathway

Aim:In our previous work,we screened the long noncoding RNA(lncRNA)expression profile of the mouse fibrotic peritoneum by lncRNA microarray.We found lncRNA-AV310809 may play a key role via Wnt2/β-catenin signaling pathway in the process of peritoneal fibrosis.In this study,our aim is to identify the effect of lncRNA-AV310809 on peritoneal fibrosis,and then explore the mechanism in HMrSV5 cells.Methods:1.HMrSV5 cells were exposed to 7ng/ml TGF-β1 for 48 h.The protein expression of FN,COL-I,E-cadherin and α-SMA were examined by Western blot;The expression level of lncRNA-AV310809 in TGF-β1-induced EMT was detected by real-time PCR;The activity of Wnt2/β-catenin signaling pathway was evaluated by Western blot.2.HMrSV5 cells were transfected with overexpressed lncRNA-AV310809 or negative control plasmid and then treated with TGF-β1.The microscopy was used to observe the phenotypic changes;Western-blot and immunofluorescence were used to examine the protein expression of FN,COL-I,E-Cadherin,α-SMA and detect the influence on Wnt2/β-catenin signaling pathway;3.MTT and the wounding heal assay were used to show the effect of lncRNA-AV310809 on the proliferation and migration in HMrSV5 cells.Results:1.Stimulation of HMrSV5 cells with TGF-β1 resulted in the decreased protein expression of E-cadherin but increased protein expression of FN,COL-I and α-SMA.2.TGF-β1 induced EMT in HMrSV5 cells with the up-regulated expression level of lncRNA-AV310809(1.30-folder,P = 0.02);3.The Wnt2/β-catenin signaling pathway was activated in the process of TGF-β1-induced EMT in HMrSV5 cells: the protein expression of Wnt2 and β-catenin were up-regulated;4.Overexpression of lncRNA-AV310809 exerted TGF-β1-induced EMT perhaps via Wnt2/β-catenin signaling pathway in HMrSV5 cells: ○1 Overexpression of lncRNA-AV310809 promoted characteristic of elongated,spindle-shaped morphology in HMrSV5 cells induced by TGF-β1;○2 Overexpression of lncRNA-AV310809 promoted the down-regulated protein expression of E-cadherin but up-regulated protein expression of FN,COL-I,and α-SMA induced by TGF-β1 in HMrSV5 cells;○3 Overexpression of lncRNA-AV310809 facilitated the up-regulated protein expression of Wnt2 and β-catenin induced by TGF-β1 in HMrSV5 cells;5.lncRNA-AV310809 may suppress the proliferation and migration in HMrSV5 cells.Conclusion:The expression level of lncRNA-AV310809 was upregulated in the process of EMT induced by TGF-β1 in HMrSV5 cells.Overexpression of lncRNA-AV310809 promoted TGF-β1-induced EMT perhaps via Wnt2/β-catenin signaling pathway in HMrSV5 cells.