摘要
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 the brain glioma stem cells originated from U87MG cells, and to study the influence of Topo- Ⅱα gene to proliferation and apoptosis of brain glioma stem cells. Methods The U87MG cells were cuhured and brain glioma stem cells were isolated and identified. Specific siRNA for Topo-Ⅱα geue was transfected into brain glioma stem cells. After transfection, the expression of Topo -Ⅱα protein was detected by Western blot, the apoptosis of the transfected cells was detected by flow cytometry. Results Neurosphere cells were isolated from U87MG cells and proved the characteristics of brain glioma stem cells. After transfection, the expression level of Topo-Ⅱα protein of the transfected brain glioma stem cells was downregulated significantly, the apoptosis rate was increased significantly comparing to the control cells. Conclusion Brain glioma stem cells were isolated successfully from U87MG cells, siRNA for Topo-Ⅱα gene can inhibit the proliferation of the ceils and increase the apoptosis of the ceils.
出处
《解剖科学进展》
CAS
2013年第5期420-423,共4页
Progress of Anatomical Sciences
基金
国家自然科学基金资助项目(No.81172197
81072056
81172408
30700861)
