转染TRIM29-siRNA对胶质瘤U87MG细胞周期和凋亡的影响

doi:10.13481/j.1671-587x.20200219

摘要/Abstract

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，G₀/G₁期、S期、G₂/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G₁/G₀期、S期、G₂/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G₂/M期U87MG细胞百分率均明显降低（P<0.05），G₀/G₁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。

关键词: 胶质瘤, 三结构域蛋白29, 细胞周期, 细胞凋亡

Abstract: Objective: To investigate the effects of tripartitemotif 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 G₀/G₁, S, G₂/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 G₀/G₁ phase, S phase,and G₂/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 G₀/G₁ 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.

Key words: glioma, tripartitemotif containing 29, cell cycle, apoptosis

中图分类号:

R739.41

肖华, 姚声涛. 转染TRIM29-siRNA对胶质瘤U87MG细胞周期和凋亡的影响[J]. 吉林大学学报(医学版), 2020, 46(02): 323-328.

XIAO Hua, YAO Shengtao. Effects of TRIM29-siRNA transfection on cell cycle and apoptosis of glioma U87MG cells[J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 323-328.

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