PEX基因修饰骨髓间质干细胞对C6胶质瘤细胞增殖和凋亡的影响  

Effect of PEX gene-modified mesenchymal stem cells on glioma cell proliferation and apoptosis

作　　者：高歌[1] 牛朝诗[2] 张俊[1] 董永飞[1] 李明武[1] 丁宛海[1] 

机构地区：[1]安徽医科大学附属省立医院神经外科,合肥230001 [2]安徽省立体定向神经外科研究所

出　　处：《中国微侵袭神经外科杂志》2010年第4期175-178,共4页Chinese Journal of Minimally Invasive Neurosurgery

基　　金：国家自然科学基金项目(编号:30672166)

摘　　要：目的探讨PEX基因修饰的骨髓间质干细胞(mesenchymal stem cells,MSC)对C6胶质瘤细胞的作用及机制。方法分子克隆技术构建PEX基因真核表达载体并转染MSC,G418筛选获取稳定表达PEX基因的MSC(MSC-PEX),将不同数量的MSC-PEX细胞与C6胶质瘤细胞进行共培养试验,用水溶性四氮唑法(WST-1)观察MSC-PEX对C6胶质瘤细胞增殖的影响,用Annexin-Ⅴ-FITC/PI双染荧光观察C6胶质瘤细胞凋亡的形态学变化,并用Annexin-Ⅴ-FITC/PI双标记法通过流式细胞仪检测胶质瘤细胞凋亡率。结果成功获得稳定表达PEX基因的MSC-PEX,MSC-PEX抑制C6胶质瘤细胞的生长作用较明显,Annexin-Ⅴ-FITC/PI双染荧光发现C6胶质瘤细胞发生凋亡形态改变;流式细胞仪检测显示:MSC-PEX转染组和DMEM对照组的凋亡率分别为16.7%和1.3%,差异有统计学意义(P<0.05)。结论PEX基因修饰的骨髓间质干细胞抑制胶质瘤细胞增殖并诱导其凋亡,为胶质瘤的治疗奠定理论基础。Objective To study the effect of PEX gene-modified mesenchymal stem cells （MSC） on C6 glioma cells in vitro and explore its mechanism.Methods PEX eukaryotic expression vector was constructed by molecular cloning and transfected MSC,then the MSC stably expressing PEX gene （MSC-PEX） were obtained by G418 selection.Variable numbers of MSC-PEX were added to C6 glioma cells by coculture experiments.Effects of MSC-PEX on C6 glioma cell growth were observed by water soluble tetrazolium （WST-1） cell proliferation and cytotoxicity assay kit.Morphological changes of C6 glioma cell apoptosis were observed by Annexin-Ⅴ-FITC and propidium iodide （Annexin-Ⅴ-FITC/PI） staining.Apoptosis rate was detected by Annexin-Ⅴ-FITC/PI dual staining assay.Results MSC-PEX was constructed successfully.MSC-PEX could inhibit the growth of C6 glioma cell line remarkably.Significant morphological changes in C6 glioma cells were observed by Annexin-Ⅴ-FITC/PI staining.The apoptosis rates of MSC-PEX transfection group and DMEM control group were 16.7% and 1.3% respectively,indicating a significant difference （P 0.05）.Conclusions PEX gene-modified mesenchymal stem cells inhibit glioma cell proliferation and induce its apoptosis,which lay a theoretical foundation for the treatment of glioma.

关 键 词：神经胶质瘤 PEX基因 间质干细胞 骨髓 

分 类 号：R73-3[医药卫生—肿瘤]