LncRNA-SNHG16 Regulates EMT Of Peritoneal Mesothelial Cells To Promote Peritoneal Fibrosis

Objective:Peritoneal dialysis(PD)has advantages over hemodialysis in terms of protecting residual renal function and allowing home treatment.However,long-term peritoneal dialysis can cause the development of peritoneal fibrosis,leading to the withdrawal of patients from PD.In this study,vitro experiments were conducted to investigate the role of long non-coding RNA small nucleolus host gene 16 in the process of peritoneal fibrosis and to provide new targets for the prevention and treatment of peritoneal fibrosis.Methods:1.Transforming growth factor β1(TGF-β1)(0,5ug/ml,7ug/ml,10ug/ml)was used to stimulate peritoneal mesothelial cell line(HMrSV5)cells for 48 h,and Western blot was used to detect the changes of E-cadherin and vimentin expression at different stimulation concentrations to determine the establishment of the stimulation concentrations for the establishment of EMT model in peritoneal mesothelial cells were determined by Western blot.2.Real-tine PCR was used to detect the changes of lncRNA-SNHG16 expression level in EMT model.FISH was performed to detect the subcellular localization of lncRNA-SNHG16 in HMrSV5 cells.3.LncRNA-SNHG16 interference vector and overexpression vector were constructed,and the expression changes of EMT-related proteins: E-cadherin,vimentin,type I collagen(COL-Ⅰ)and fibronectin(FN)were detected by Western blot after the interference and overexpression of SNHG16.The expression changes and localization of EMT-associated proteins: E-cadherin and vimentin in HMRSV5 cells were detected by IF.Results:1.Stimulation with TGF-β1(10ug/ml,48h)can induce in vitro peritoneal mesothelial cell EMT.2.In the induction of EMT in HMrSV5 cells by TGF-β1,Real-time PCR detected up-regulated expression of lncRNA-SNHG16.FISH showed that lncRN A-SNHG16 was expressed in both cytoplasm and nucleus of HMrSV5 cells.3.Overexpression of lncRNA-SNHG16 promoted TGF-β1-induced EMT in HMrSV5 cells in vitro: EMT-associated epithelial phenotype E-cadherin expression decreased,while mesenchymal phenotype protein vimentin,COL-Ⅰ,and fibronectin expression increased.4.Interference with lncRNA-SNHG16 inhibited TGF-β1-induced peritoneal mesothelial EMT in vitro: expression of epithelial phenotype protein E-cadherin was increased,and expression of mesenchymal phenotype protein vimentin,COL-Ⅰ,and fibronectin expression was decreased.Conclusion:LncRNA-SNHG16 expression was upregulated in the EMT model of HMrSV5 cells,and in vitro experiments verified that it promoted TGF-β1-induced EMT occurrence in HMrSV5 cells and was involved in peritoneal fibrosis in peritoneal mesothelial cells.