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目的由U87MG细胞分离并鉴定脑胶质瘤干细胞,研究Topo-Ⅱα基因对脑胶质瘤干细胞增殖和凋亡的影响。方法培养脑胶质瘤U87MG细胞,分离和鉴定其中的胶质瘤干细胞。将Topo-Ⅱα基因特异性的siRNA转染胶质瘤干细胞。转染后,Western Blot检测转染后Topo-Ⅱα蛋白的表达,MTT法检测细胞增殖活力,双标流式法检测细胞凋亡率。结果成功分离出U87MG细胞球细胞,并鉴定其具有肿瘤干细胞特性。转染Topo-Ⅱα基因特异性的siRNA后,胶质瘤干细胞中的Topo-Ⅱα蛋白表达明显降低,同时,细胞增殖能力显著降低,凋亡率明显增加。结论由U87MG细胞系中分离出细胞球细胞具备肿瘤干细胞特性,Topo-Ⅱα基因沉默降低胶质瘤干细胞的增殖能力,促进细胞凋亡。
Objective To isolate and identify glioma stem cells from U87MG cells and study the effect of Topo-Ⅱα gene on the proliferation and apoptosis of glioma stem cells. Methods Glioma U87MG cells were cultured and the glioma stem cells were isolated and identified. Topo-IIα gene-specific siRNA was transfected into glioma stem cells. After transfection, the expression of Topo-Ⅱαprotein was detected by Western Blot, the cell proliferation activity was detected by MTT assay, and the apoptosis rate was detected by double-labeled flow cytometry. Results U87MG cells were successfully isolated and identified as tumor stem cells. Topo-Ⅱα gene-specific siRNA transfection, Topo-Ⅱ α protein expression in glioma stem cells was significantly reduced, while significantly reduced cell proliferation, apoptosis rate was significantly increased. Conclusion The characteristics of tumor stem cells isolated from U87MG cell line are tumor stem cells. Topo-Ⅱ α gene silencing can reduce the proliferation of glioma stem cells and promote apoptosis.