miR-486对CD133+胶质瘤干细胞的影响

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，实时定量PCR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 胶质瘤干细胞纯度达到实验要求，CD133 胶质瘤细胞中miR-486表达量较CD133-胶质瘤细胞中下降（P<0.01），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，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（P<0.05），转染miR-486后细胞凋亡率显著提高（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中队胶质瘤干细胞的有效靶点。

Influence of miR-486 on CD133+ glioma stem cells

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.