Verification And Functional Study Of Lnc-PCK1-2:1 In Epithelial-mesenchymal Transition Of Human Lens Epithelial Cells

Objective: To investigate the expression of lnc-PCK1-2:1 in epithelial-mesenchymal transition of human lens epithelial cells and to explore its molecular function.Methods: Epithelial-mesenchymal transdifferentiation was induced in human lens epithelial cells by applying 10 ng/ml of TGF-β2 in the early stage.Then,differentially expressed lncRNAs were screened by gene chip technology,and the top 10 lncRNAs were screened.In this experiment,one of the lncRNAs-lnc-PCK1-2:1 was selected for validation and functional studies.qRT-PCR was used to verify the expression of lnc-PCK1-2:1 during epithelial-mesenchymal differentiation of human lens epithelial cells.The siRNA-lnc-PCK1-2:1 plasmid was transfected with 10 ng/ml TGF-β2 in DMEM medium,which was the siRNA-lnc-PCK1-2:1+ TGF-β2 group.10 ng/ml TGF-β2 in DMEM medium,and then transfected with siRNA-NC plasmid,this is siRNA-NC+ TGF-β2 group.Only the DMEM medium was added,and the siRNA-lnc-PCK1-2:1 plasmid was transfected,which was siRNA-lnc-PCK1-2:1group.Only DMEM medium was added and the siRNA-NC plasmid was transfected,which was the siRNA-NC group.Cellular immunofluorescence and Western blot were used to observe the expression of EMT-related moleculessmooth muscle actin(a-SMA)and E-cadherin(E-Cadherin)at the protein level.The cells were detected by qRT-PCR.Changes in mRNA expression.Results: It was confirmed that lnc-PCK1-2:1 was expressed during epithelial-mesenchymal transition in human lens epithelial cells.In the siRNA-lnc-PCK1-2:1+ TGF-β2group,siRNA-NC+ TGF-β2 group,the fluorescence intensity of a-SMA increased gradually,protein expression increased gradually;E-Cadherin fluorescence intensity gradually weakened,protein The expression decreased gradually.In the siRNA-lnc-PCK1-2:1 group and siRNA-NC group,the fluorescence intensity of a-SMA increased gradually and the protein expression gradually increased.The fluorescence intensity of E-Cadherin gradually decreased,and the protein expression gradually decreased.The fluorescence intensity and protein expression of a-SMA were higher in siRNA-lnc-PCK1-2:1+TGF-β2 group than in siRNA-lnc-PCK1-2:1 group,siRNA-NC+ TGF-β2 group was larger than siRNA-NC group,E-Cadherin fluorescence intensity,protein expression siRNA-lnc-PCK1-2:1+TGF-β2 group is smaller than siRNA-lnc-PCK1-2:1 group,siRNA-NC+ TGF-β2 group is smaller than siRNA-NC group;The expression level of lnc-PCK1-2:1 was greater in siRNA-lnc-PCK1-2:1+TGF-β2 group than in siRNA-NC+ TGF-β2group,siRNA-lnc-PCK1-2:1 group was larger than siRNA-NC group,siRNA-lnc-PCK1-2:1+ TGF-β2 group is greater than siRNA-lnc-PCK1-2:1 group,siRNA-NC+ TGF-β2 group is larger than siRNA-NC group.Conclusion:Lnc-PCK1-2:1 is highly expressed during epithelial-mesenchymal transition in human lens epithelial cells and silencing of lnc-PCK1-2:1 inhibits epithelial-mesenchymal transition in lens epithelial cells.