首页|期刊导航|山东医药|lncRNA-MEG3敲低人子宫平滑肌瘤细胞株增殖凋亡、克隆形成、细胞周期变化及与miR-93靶向关系

lncRNA-MEG3敲低人子宫平滑肌瘤细胞株增殖凋亡、克隆形成、细胞周期变化及与miR-93靶向关系OACSTPCD

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

中文摘要英文摘要

目的 观察敲低长链非编码核糖核酸-MEG3(lncRNA-MEG3)对人子宫平滑肌瘤(UL)细胞株的增殖凋亡、克隆形成、细胞周期的影响,并分析lncRNA-MEG3与微小核糖核酸-93(miR-93)的靶向关系.方法 体外培养人UL细胞株SK-UT-1,细胞分为lncRNA-MEG3敲低组和敲低阴性对照组,分别转染lncRNA-MEG3敲低质粒和敲低阴性对照质粒至两组细胞,采用qRT-PCR法检测细胞中lncRNA-MEG3和miR-93,CCK8法检测细胞增殖活性,平板克隆形成实验检测细胞克隆形成能力,流式细胞术检测细胞周期及凋亡率,Western blotting法检测增殖相关蛋白(Ki67、PCNA蛋白)、凋亡相关蛋白(Bax、Bcl-2蛋白)及Wnt/β-catenin通路相关蛋白(Wnt3a、β-catenin蛋白).采用双荧光素酶报告基因实验验证lncRNA-MEG3和miR-93的靶向关系.结果 与敲低阴性对照组比较,lncRNA-MEG3敲低组细胞lncRNA-MEG3表达降低,miR-93表达升高,48、72、96 h时OD值降低,克隆形成数目减少,G0/G1期占比增加,G2/M期占比下降,凋亡率升高,Wnt3a、β-catenin、Ki67、PCNA、Bcl-2蛋白表达降低,Bax蛋白表达升高(P均<0.05).lncRNA-MEG3能靶向miR-93从而降低相对荧光素酶活性(P<0.05).结论 敲低lncRNA-MEG3能够抑制SK-UT-1细胞的增殖、克隆形成及Wnt/β-catenin通路,将细胞周期阻滞于G0/G1期,并诱导细胞凋亡,从而抑制细胞生长,lncRNA-MEG3与miR-93存在靶向关系.

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.

马文文;曹晖;陈茜松;于倩;冬国友

唐山市妇幼保健院妇产科,河北唐山 063000唐山市妇幼保健院病理科

临床医学

子宫平滑肌瘤微小核糖核酸-93细胞增殖细胞凋亡细胞克隆形成细胞周期Wnt/β-catenin通路长链非编码核糖核酸-MEG3

uterine leiomyomamicroRNA-93cell proliferationapoptosisclone formationcell cycleWnt/β-catenin pathwaylong non-coding RNA-MEG3

《山东医药》 2024 (036)

1-5 / 5

河北省医学研究课题计划(20221755).

10.3969/j.issn.1002-266X.2024.36.001

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