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  

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

作　　者：马文文 曹晖 陈茜松[1] 于倩 冬国友[2] MA Wenwen;CAO Hui;CHEN Qiansong;YU Qian;DONG Guoyou(Department of Obstetrics and Gynecology,Tangshan Maternal and Child Health Hospital,Tangshan 063000,China;不详)

机构地区：[1]唐山市妇幼保健院妇产科,河北唐山063000 [2]唐山市妇幼保健院病理科

出　　处：《山东医药》2024年第36期1-5,共5页Shandong Medical Journal

基　　金：河北省医学研究课题计划(20221755)。

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,G_(0)/G_(1)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G_(0)/G_(1)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,clonal 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 detected by plate colony formation assay,the cell cycle and apoptosis rate were detected by flow cytometry,and the expression 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 lncRNA-MEG3 and miR-93 was verified by dual luciferase reporter gene assay.Results Compared with the negative control 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 G_(0)/G_(1) 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 G_(0)/G_(1) phase,and induce apoptosis,thereby inhibiting the growth of UL cells.There is a targeted relationship between lncRNA-MEG3 and miR-93.

关 键 词：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 

分 类 号：R737[医药卫生—肿瘤]