转染TRIM29-siRNA对胶质瘤U87MG细胞周期和凋亡的影响  

Effects of TRIM29-siRNA transfection on cell cycle and apoptosis of glioma U87MG cells

作　　者：肖华 姚声涛 XIAO Hua;YAO Shengtao(Department of Cerebrovascular Diseases,Affiliated Hospital,Zunyi Medical University,Zunyi 563099,China)

机构地区：[1]遵义医科大学附属医院脑血管病科,贵州遵义563099

出　　处：《吉林大学学报（医学版）》2020年第2期323-328,共6页Journal of Jilin University：Medicine Edition

基　　金：贵州省科技厅科技计划项目资助课题(黔科合平台人才〔2018〕3056号);贵州省遵义市科技局科研基金资助课题(遵市科合HZ字〔2019〕65号)。

摘　　要：目的:探讨转染三结构域蛋白(TRIM)家族成员三结构域蛋白29(TRIM29)-siRNA对胶质瘤U87MG细胞周期和凋亡的影响,阐明TRIM29在胶质瘤发生发展中的作用。方法:将体外培养的U87MG细胞分为对照组(未转染)、NC-siRNA组(转染阴性对照NC-siRNA)和TRIM29-siRNA组(转染TRIM29-siRNA),采用实时荧光定量PCR法检测3组U87MG细胞中TRIM29 mRNA表达水平,流式细胞术检测3组不同细胞周期U87MG细胞百分率和凋亡率,Western blotting法检测3组细胞中TRIM29、CyclinD1、P21、Bcl-2和Bax蛋白表达水平。结果:3组细胞中TRIM29、CyclinD1、P21、Bcl-2和Bax蛋白表达水平,TRIM29 mRNA表达水平,G 0/G 1期、S期、G 2/M期U87MG细胞百分率及细胞凋亡率比较差异有统计学意义(P<0.05)。与对照组比较,NC-siRNA组U87MG细胞中TRIM29 mRNA和TRIM29、CyclinD1、Bcl-2蛋白表达水平和G 1/G 0期、S期、G 2/M期U87MG细胞百分率及细胞凋亡率差异无统计学意义(P>0.05);与对照组和NC-siRNA组比较,TRIM29-siRNA组U87MG细胞中TRIM29 mRNA和TRIM29、CyclinD1、Bcl-2蛋白表达水平以及S期和G 2/M期U87MG细胞百分率均明显降低(P<0.05),G 0/G 1期U87MG细胞百分率和细胞凋亡率以及P21、Bax蛋白表达水平明显升高(P<0.05)。结论:转染TRIM29-siRNA可诱导U87MG细胞周期阻滞和凋亡,其作用机制可能与下调CyclinD1、Bcl-2蛋白表达和上调P21、Bax蛋白表达有关。Objective:To investigate the effects of tripartite motif containing 29(TRIM29)-siRNA transfection on the cell cycle and apoptosis of glioma U87MG cells,and to elucidate the role of TRIM29 in the occurrence and development of glioma.Methods:The U87MG cells cultured in vitro were divided into control group(untransfected),NC-siRNA group(transfected with negative control NC-siRNA),and TRIM29-siRNA group(transfected with TRIM29-siRNA).The expression levels of TRIM29 mRNA in the U87MG cells in three groups were detected by real-time fluorescent quantitative PCR,the percentages of U87MG cells in different cell cycles and the apoptotic rates in three groups were detected by flow cytometry,and the expressions levels of TRIM29,CyclinD1,p21,Bcl-2 and Bax proteins in the U87MG cells in three groups were detected Western blotting method.Results:There were significant differences in the expression levels of TRIM29,CyclinD1,P21,Bcl-2,Bax proteins,the expression levels of TRIM29 mRNA,and the percentages of U87MG cells in G 0/G 1,S,G 2/M phases and the apoptotic rates among three groups(P<0.05).Compared with control group,the expression levels of TRIM29 mRNA and TRIM29,CyclinD1,Bcl-2 proteins,the percentages of U87MG cells in G 0/G 1 phase,S phase,and G 2/M phase,and the apoptotic rates in the U87MG cells in NC-siRNA group had no significant differences(P>0.05).Compared with control group or NC-siRNA group,the expression levels of TRIM29 mRNA and TRIM29,CyclinD1,Bcl-2 proteins in the U87MG cells in TRIM29-siRNA group were decreased significantly(P<0.05);the percentage of U87MG cells in G 0/G 1 phase,the apoptotic rate of U87MG cells and the expression levels of p21 and Bax proteins were increased significantly(P<0.05).Conclusion:TRIM29-siRNA transfection can induce the cell cycle arrest and apoptosis in the U87MG cells and its mechanism may be related to down-regulation of Cyclin D1,Bcl-2 protein expressions and up-regulation of p21 and Bax protein expressions.

关 键 词：胶质瘤 三结构域蛋白29 细胞周期 细胞凋亡 

分 类 号：R739.41[医药卫生—肿瘤]