| 韩福新,张蕊,马善波,等.miR-486对CD133+胶质瘤干细胞的影响[J].安徽医药,2016,20(12):2323-2326. |
| miR-486对CD133+胶质瘤干细胞的影响 |
| Influence of miR-486 on CD133+ glioma stem cells |
| 投稿时间：2016-03-14 |
| DOI： |
| 中文关键词: 胶质瘤干细胞 miR-486 细胞增殖 |
| 英文关键词: Glioma stem cell miR-486 Proliferation |
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| 中文摘要: |
| 目的 研究miR-486对CD133+胶质瘤干细胞的影响。方法 CD133抗体标记U87胶质瘤细胞系,流式细胞分选纯化CD133+胶质瘤干细胞,实时荧光定量PCR(qPCR)检测CD133+胶质瘤干细胞中miR-486的表达量。脂质体转染方法构建过表达miR-486的CD133+胶质瘤干细胞,MTT法检测其凋亡率,流式分析仪检测细胞增殖率与生长周期。结果 流式分选后CD133+胶质瘤干细胞纯度从1.7%提升至84.2%(P<0.001),达到实验要求。CD133+U87胶质瘤细胞中miR-486相对表达量为(0.33±0.10),显著低于CD133+U87胶质瘤细胞中的miR-486表达量(0.45±0.09)(P<0.05)。转染miR-486模拟物的CD133+胶质瘤干细胞较转染miRNA对照模拟物的CD133+胶质瘤干细胞相对增殖率降低(220% vs 360%),细胞周期检测对照组细胞位于G0/G1期的比例低于转染miR-486后的细胞(62.8% vs 76.8%,P<0.05),转染miR-486后细胞凋亡率显著提高(1.7% vs 23.2%,P<0.001)。结论 CD133+胶质瘤干细胞中miR-486表达量减少,过表达miR-486的CD133+胶质瘤干细胞增殖受到抑制,生长周期被阻滞于G1/S期,凋亡率显著提高,可能是胶质瘤靶向治疗胶质瘤干细胞的有效靶点。 |
| 英文摘要: |
| Objective To study the influence of miR-486 on CD133+ glioma stem cells.Methods We labeled U87 glioma cell line by antibody of CD133,separated and purified CD133+ glioma stem cells by flow cytometry,and detected the expression of miR-486 in CD133+ glioma stem cells by real-time quantitative polymerase chain reaction (qPCR) .We then constructed CD133+ glioma stem cells with over-expressed miR-486 by liposome transfection,and then tested the proliferation by MTT and analyzed apoptosis rate and growth cycle by flow cytometry.Results After streaming sorting,the purity of CD133+ glioma stem cells increased from 1.7% to 84.2% (P<0.001),which met the experimental requirements.The miR-486 relative expression of CD133+ U87 glioma cells was (0.33±0.10),significantly lower than that of CD133+ U87 glioma cells (0.45±0.09,P<0.05).The relative proliferation rate of the CD133+ glioma stem cell with transfected miR-486 simulation was lower than that with transfected miRNA simulation (220% vs 360%).The cell proportion of the control group in G0/G1 phase was lower than the cell with overexpressed miR-486 (62.8% vs 76.8%,P<0.05).The apoptosis rate of the miR-486 overexpression cell increased significantly (1.7% vs 23.2%,P<0.001).Conclusions The expression of miR-486 decreased in CD133+ glioma stem cells.The over-express of miR-486 glioma cells restrained proliferation,blocked the cell cycle in G1/S suppressed proliferation of CD133+ glioma stem cells,the restrained G1/S phase,and the significantly increased apoptosis may be the effective target for targeted therapy of glioma. |
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