微小核糖核酸592对神经胶质瘤细胞株U251细胞凋亡的影响

Effect of microRNA-592 on the apoptosis of glioma cell line U251

摘要:

目的 研究微小核糖核酸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，并对U251细胞的凋亡进行分析。结果 对28份神经胶质瘤组织和正常组织的定量PCR结果分析发现，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；流式细胞分析结果显示，miR-592显著促进U251细胞凋亡：实验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结果发现miR-592直接靶向Runx2的3'-UTR来抑制Runx2蛋白的水平；接下来，向U251细胞转染Runx2的siRNA，绘制细胞的生长曲线，并通过流式细胞技术分析U251细胞的凋亡率。结论 miR-592通过直接靶向Runx2来诱导神经胶质瘤细胞凋亡，进而抑制肿瘤细胞的生长。

Abstract:

Objective To investigate the effect of microRNA-592(miR-592) on the apoptosis of glioma cell line U251.Methods PCR was used to measure the expression of miR-592 in glioma tissue and adjacent tissue.U251 cells were transfected with miR-592 mimics and then flow cytometry was used to analyze the effect of miR-592 overexpression on the apoptosis of U251 cells.A bioinformatics analysis was used to identify the potential target molecules of miR-592,and then dual-luciferase reporter assay and Western blot were used for validation.U251 cells were transfected with the small interfering RNA (siRNA) which downregulated the expression of Runx2,and then the cell growth curve was plotted and the apoptosis of U251 cells was analyzed.Results The results of quantitative PCR for 28 glioma tissues and normal tissues showed that the expression of miR-592 was significantly reduced in glioma tissues and miR-592 overexpression significantly inhibited the growth of U251 cells.Flow cytometry showed that miR-592 significantly promoted the apoptosis of U251 cells;the late apoptosis rate was 7.2%±0.68% in the control group and 17.47%±1.45% in the miR-592 transfection group.Dual-luciferase reporter assay and Western blot showed that miR-592 directly targeted 3'-UTR of Runx2 to inhibit the level of Runx2 protein.U251 cells were transfected with the siRNA of Runx2;the cell growth curve was plotted,and flow cytometry was used to analyze the apoptosis rate of U251 cells.Conclusions miR-592 directly targets Runx2 to induce the apoptosis of U251 cells and thus inhibit cell growth.