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目的探讨微小核糖核酸592(miR-592)对神经胶质瘤细胞株U251凋亡的影响。方法首先通过定量聚合酶链反应(PCR)分析miR-592在28份神经胶质瘤与其临近癌旁组织中的表达水平;随后向U251细胞转染miR-592的拟合物,并通过流式细胞技术分析miR-592过表达对U251细胞凋亡的影响;通过生物信息学分析找到miR-592的潜在靶分子,并通过荧光素酶双报告实验以及蛋白免疫印迹法等进行验证;进一步转染U251细胞Runx2的下调siRNA,绘制细胞的生长曲线,检测U251细胞的凋亡率。结果对28份神经胶质瘤组织和正常组织的定量PCR分析发现,miR-592在肿瘤组织中明显低表达;miR-592过表达能明显抑制U251细胞的生长;流式细胞分析显示,miR-592显著促进U251细胞凋亡:对照组晚期凋亡率为(7.2±0.68)%,而转染miR-592组晚期凋亡率为(17.47±1.45)%;荧光素酶双报告实验以及蛋白免疫印迹法实验发现miR-592直接靶向Runx2的3’-UTR来抑制Runx2蛋白的表达水平。结论 miR-592通过直接靶向Runx2来诱导神经胶质瘤细胞凋亡,进而抑制细胞生长。
Objective To investigate the effect of miRNA 592 (miR-592) on the apoptosis of glioma cell line U251. Methods The expression level of miR-592 in 28 gliomas and adjacent normal tissues was detected by quantitative polymerase chain reaction (PCR). The miR-592 was then transfected into U251 cells, Cell technology analysis of miR-592 overexpression on U251 cell apoptosis; By bioinformatics analysis to find potential targets of miR-592 molecules, and through luciferase dual-reporter experiments and Western blot analysis to verify; further transfection U251 cells Runx2 down-regulation of siRNA, the growth curve of the cells were drawn to detect the apoptosis rate of U251 cells. Results Quantitative PCR analysis of 28 glioma tissues and normal tissues showed that miR-592 was significantly down-regulated in tumor tissues. Overexpression of miR-592 significantly inhibited the growth of U251 cells. Flow cytometry analysis showed that miR- 592 significantly promoted the apoptosis of U251 cells: the late apoptotic rate was (7.2 ± 0.68)% in the control group and (17.47 ± 1.45)% in the transfected miR-592 group; the double luciferase reporter assay and the protein immunization Western blotting showed miR-592 directly targeted Runx2 3’-UTR to suppress Runx2 expression. Conclusion miR-592 induces apoptosis of glioma cells by directly targeting Runx2, thereby inhibiting cell growth.