Effect of lncRNA TP73-AS1 on ovarian granulosa cell apoptosis in polycystic ovary syndrome by regulating miR-128-3p
Objective To investigate the role of lncRNA TP73-AS1 in polycystic ovary syndrome and its potential mechanism.Methods Human ovarian granulosa cell line KGN cells were cultured in vitro,and Vector,pcDNA-TP73-AS1,NC-siRNA,TP73-AS 1-siRNA,TP73-AS 1-siRNA+NC inhibitor,TP73-AS1-siRNA+miR-128-3p inhibitor were transfected into KGN cells,respectively.Real-time quantitative PCR(RT-qPCR)experiments were used to detect the expression levels of lncRNA TP73-AS1 and miR-128-3p.MTT assay was used to detect cell proliferation.The apoptosis rate was detected by flow cytometry.Bioinformatics software and dual luciferase reporter assay were used to predict and verify the targeting relationship between lncRNA TP73-AS1 and miR-128-3p.Results After cells were transfected with pcDNA-TP73-AS1,the expression of lncRNA TP73-AS1 was up-regulated,the cell proliferation ability was increased,and the apoptosis rate was decreased(P＜0.05).After cells were transfected with TP73-AS 1-siRNA,the expression of lncRNA TP73-AS1 was down-regulated,the cell proliferation ability was decreased,and the apoptosis rate was increased(P＜0.05).Dual luciferase reporter assay showed that miR-128-3p was the target gene of lncRNA TP73-AS1.The expression of miR-128-3p in TP73-AS1-siRNA group was significantly higher than that in NC-siRNA group(P＜0.05).The expression of miR-128-3p in pcDNA-TP73-AS1 group was significantly lower than that in Vector group(P＜0.05).miR-128-3p inhibitor treatment could significantly reverse the regulatory effects of TP73-AS1-siRNA on cell proliferation and apoptosis.Conclusion Knockdown of lncRNA TP73-AS1 could inhibit ovarian granulosa cell proliferation and promote apoptosis by targeting miR-128-3p.
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