miR-186对胶质母细胞瘤细胞增殖和凋亡的影响及机制

发布时间：2018-03-18 04:25

  本文选题：miR-　切入点：胶质母细胞瘤　出处：《华中科技大学学报(医学版)》2017年06期 　论文类型：期刊论文

【摘要】：目的研究转染miR-186对胶质母细胞瘤细胞增殖能力的影响并探讨作用机制。方法采用荧光定量PCR检测miR-186在胶质母细胞瘤细胞系U251、U87-MG及正常星形胶质细胞系HA1800中的表达水平,将U251细胞分为miR-186模拟物组和对照组,分别转染miR-186模拟物和阴性随机对照序列,并用荧光定量PCR检测miR-186在两组中的表达量。采用CKK-8法检测两组细胞细胞增殖能力,流式细胞仪测定两组细胞凋亡率,Western blot分析Caspase-3、Caspase-8、Caspase-9及Cyclin D1蛋白的表达。结果 miR-186低表达于胶质母细胞瘤细胞系U251及U87-MG,高表达于正常星形胶质细胞系HA1800。转染24、48、72及96h后,miR-186模拟物组细胞增殖(A450nm值)低于对照组(均P0.05)。转染48h后,miR-186模拟物组凋亡率高于对照组;miR-186模拟物组Caspase-3、9的表达量高于对照组(均P0.01),Caspase-8与对照组相比差异无统计学意义(P0.05)。miR-186模拟物组Cyclin D1表达水平较对照组明显降低(P0.01)。结论miR-186抑制胶质母细胞瘤细胞增殖能力,并诱导凋亡,其机制可能与上调Caspase-3、9,下调Cyclin D1有关。
[Abstract]:Objective to investigate the effect of miR-186 transfection on the proliferation of glioblastoma cells and its mechanism. Methods the expression of miR-186 in glioblastoma cell line U251 U87-MG and normal astrocytoma cell line HA1800 was detected by fluorescence quantitative PCR. U251 cells were divided into miR-186 mimics group and control group. MiR-186 mimics and negative random control sequences were transfected into U251 cells respectively. The expression of miR-186 in the two groups was detected by fluorescence quantitative PCR. The proliferation ability of the two groups was detected by CKK-8 assay. The expression of Caspase-3, Caspase-8, Caspase-9 and Cyclin D1 protein was detected by flow cytometry. Results the expression of miR-186 was low in glioblastoma cell lines U251 and U87-MG, and highly expressed in normal astrocytoma cell line HA1800.After transfection, the miR-186 model was obtained. After 48 hours of transfection, the apoptotic rate of the mimetic group was higher than that of the control group. The expression of Caspase-3O9 in the control group was higher than that in the control group (P 0.01). There was no significant difference in the expression of Caspase-8 between the control group and the control group (P0.05.miR-186). The expression of Cyclin D1 in the control group was significantly lower than that in the control group. Conclusion miR-186 can inhibit the proliferation of glioblastoma cells. The mechanism of apoptosis may be related to the up-regulation of Caspase-3 and down-regulation of Cyclin D1.
【作者单位】： 华中科技大学同济医学院附属武汉中心医院麻醉科;华中科技大学同济医学院附属武汉中心医院急诊创伤外科;
【分类号】：R739.41
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本文编号：1627995