微小核糖核酸592调控神经胶质瘤细胞株U251细胞凋亡

发布时间：2018-11-23 09:38

【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分析miR-592在神经胶质瘤组织和癌旁组织中的表达变化;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si RNA,绘制细胞的生长曲线,并对U251细胞的凋亡进行分析。结果定量PCR结果分析发现,miR-592在肿瘤组织中明显低表达(t=2.752,P=0.013);miR-592过表达能明显抑制U251细胞的生长,与对照组比较,培养36 h、48 h后存在统计学差异(t=2.127,P=0.031;t=2.284,P=0.026);流式细胞分析结果显示,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%,存在统计学差异(t=3.294,P=0.007);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;检测下调Runx2对U251细胞生长的影响,结果显示转染了Runx2 si RNA的细胞生长明显比对照组低(t=3.124,P=0.011),流式细胞技术对周期的检测显示,Runx2下调表达上调U251细胞的凋亡率。通过绘制肿瘤生长曲线,发现miR-592过表达明显抑制肿瘤的生长。同时,Runx2的下调表达也明显抑制肿瘤的生长。结论 miR-592通过直接靶向Runx2来诱导神经胶质瘤细胞凋亡,进而抑制细胞的生长。
[Abstract]:Objective to study the effect of microRNA 592 (miR-592) on apoptosis of glioma cell line U251. Methods the expression of miR-592 in gliomas and adjacent tissues was analyzed by quantitative polymerase chain reaction (QPCR). The miR-592 fitting was transfected into U251 cells and the effect of overexpression of miR-592 on the apoptosis of U251 cells was analyzed by flow cytometry. The potential target molecules of miR-592 were identified by bioinformatics analysis and verified by luciferase double report assay and Western blot. Furthermore, the down-regulated si RNA, of U251 cells transfected with Runx2 was used to draw the growth curve of U251 cells, and the apoptosis of U251 cells was analyzed. Results the results of quantitative PCR analysis showed that the expression of miR-592 was significantly lower in tumor tissues (tr 2.752). Overexpression of miR-592 could significantly inhibit the growth of U251 cells. Compared with the control group, there was a significant difference in the growth of U251 cells cultured for 36 h or 48 h (t = 2.127, P = 0.031, P = 2.284, P = 0.026). Flow cytometry analysis showed that miR-592 significantly promoted U251 cell apoptosis: the late apoptosis rate was 7.2% 卤0.68 in the control group and 17.47% 卤1.45% in the miR-592 transfection group. P0. 007); The results of luciferase double report assay and Western blot assay showed that miR-592 directly targeted the 3'-UTR of Runx2 to inhibit the level of Runx2 protein. The effect of down-regulation of Runx2 on the growth of U251 cells was detected. The results showed that the growth of U251 cells transfected with Runx2 si RNA was significantly lower than that of the control group (tr 3.124). Flow cytometry showed that the down-regulated expression of Runx2 upregulated the apoptosis rate of U251 cells. By drawing the tumor growth curve, it was found that overexpression of miR-592 significantly inhibited tumor growth. At the same time, the down-regulated expression of Runx2 also significantly inhibited the growth of tumor. Conclusion miR-592 can directly target Runx2 to induce glioma cell apoptosis and then inhibit the growth of glioma cells.
【作者单位】：滨州市中心医院肿瘤科;新疆医科大学第一附属医院儿外一科;
【分类号】：R739.41

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