摘要
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 miR-592 on U251 cell apoptosis in the Glioma.Methods The expression of miR-592 was analyzed by quantitative PCR in Glioma tissues from patients.Next,U251 cells were transfected with miR-592 mimics and then the growth of cells was detected by MTT assay.To extensively explore the direct target of miR-592 in glioma,dual-luciferase reporter assay and western blot assay were performed to confirm that Runx2 is the direct target of miR-592 in U251 cells.In order to test whether Runx2 was the functional target of miR-592,the cell growth curve was determined by down-regulating the level of Runx2.Moreover,the apoptosis of U251 was also detected after Runx2 knocking down.Results The expression of miR-592 was significantly reduced in glioma tissues.Over-expression miR-592 remarkably increased the apoptotic rate of U251 cells.Dual-luciferase reporter assay indicated that Runx2 was the direct target of miR-592 in U251 cells.Suppressed expression of Runx2 by siRNA prominently suppressed U251 cell growth and induced the cell apoptotic rate.Conclusion miR-592 suppressed the growth and promoted the apoptotic rate of U251 cells by targeting Runx2.
出处
《卒中与神经疾病》
2017年第4期319-322,341,共5页
Stroke and Nervous Diseases
