Effects of lncRNA-MEG3 knockdown on proliferation,apoptosis,clone formation and cell cycle,and the targeted relationship between lncRNA-MEG3 and miR-93 in human uterine leiomyoma cells
Objective To investigate the effects of lncRNA-MEG3 knockdown on the proliferation,apoptosis,clon-al formation,cell cycle,and to analyze the targeted relationship between lncRNA-MEG3 and miR-93 in human uterine leiomyoma(UL)cell line.Methods Human UL cell line SK-UT-1 was cultured in vitro,and the cells were divided into lncRNA-MEG3 knockdown group and negative control group;the lncRNA-MEG3 knockdown plasmid and negative control plasmid were transfected into cells of the two groups,respectively.The expression of lncRNA-MEG3 and miR-93 in the cells was detected by qRT-PCR,the proliferation activity was detected by CCK8 assay,the clonal formation ability was de-tected by plate colony formation assay,the cell cycle and apoptosis rate were detected by flow cytometry,and the expres-sion levels of proliferation-related proteins(Ki67,PCNA),apoptosis-related proteins(Bax,Bcl-2),and Wnt/β-catenin pathway-related proteins(Wnt3a,β-catenin)were detected by Western blotting.The targeted relationship between ln-cRNA-MEG3 and miR-93 was verified by dual luciferase reporter gene assay.Results Compared with the negative con-trol group,the expression of lncRNA-MEG3 decreased,the expression of miR-93 increased,the OD values decreased at 48,72,and 96 h,the number of clonal formation was reduced,the proportion of cells in the G0/G1 phase increased,the proportion of cells in the G2/M phase decreased,the apoptosis rate increased,the expression of Wnt3a,β-catenin,Ki67,PCNA,Bcl-2 decreased,and the expression of Bax increased in the lncRNA-MEG3 knockdown group(all P<0.05).Ln-cRNA-MEG3 could target miR-93 to reduce the relative luciferase activity(P<0.05).Conclusions Knocking down ln-cRNA-MEG3 can inhibit the proliferation,colony formation,and Wnt/β-catenin pathway of UL cells,block the cell cycle in G0/G1 phase,and induce apoptosis,thereby inhibiting the growth of UL cells.There is a targeted relationship between lncRNA-MEG3 and miR-93.
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