PEX基因对C6胶质瘤干细胞增殖作用影响的研究  

Study on effect of PEX gene-modified C6 glioma stem cells on proliferation

作　　者：黄炜[1] 张云峰[1] 高歌[2] 

机构地区：[1]海军安庆医院神经外科,246003 [2]安徽省立医院神经外科

出　　处：《临床神经外科杂志》2014年第5期326-329,334,共5页Journal of Clinical Neurosurgery

基　　金：国家自然科学基金项目(项目编号30672166)

摘　　要：目的探讨PEX基因对大鼠C6胶质瘤干细胞增殖和细胞周期的影响。方法采用传统的血清培养基培养C6胶质瘤细胞系,再取其单细胞悬液悬浮培养于无血清培养基中传代培养。采用免疫荧光染色分离鉴定脑肿瘤干细胞(BTSCs),应用本实验室成功构建的重组子pDsRed2-C1-PEX以脂质体为介导转染体外培养的BTSCs.RT-PCR验证真核表达载体在BTSCs中的表达。MTT法观察PEX对BTSCs增殖活性的影响,并用流式细胞仪检测胶质瘤干细胞的细胞周期变化。结果成功培养出BTSCs并成功转染,PEX在BTSCs中高表达并且其对体外培养的BTSCs的增殖有明显抑制作用,流式细胞仪检测细胞周期显示:BTSCsPEX转染组细胞周期G0/G1期细胞百分数明显升高为81.10%,S期细胞百分数明显降低为12.04%,差异有统计学意义(P<0.05)。结论 PEX基因可以在BTSCs中表达,并且能直接抑制体外培养的BTSCs的生长增殖。Objective To study the effects of PEX gene on proliferation and cell cycle of C 6 glioma stem cells in vitro .Methods Using traditional serum medium cultured C 6 cell lines and taking the single-celled levitation liquid suspension subculture in serum-free, brain tumor stem cells were separated and identified by immune fluorescence staining with pDsRed 2-C1-PEX which the lab successful constuction were transfected .In the vitro brain tumor stem cells （ BTSCs ） by liposomemediated,and then was detected by using reverse transcriptase-polymerase chain reaction .Effects of PEX on BTSCs growth were observed by water soluble tetrazolium cell proliferation and cytotoxicity assay kit .The cell cycle changes of BTSCs was detected by flow cytometry analysis .Results The BTSCs were cultured successfully and were transfected by PEX high expression of PEX in BTSCs in vitro and its inhibition of the proliferation of BTSCs were detected .The group which carrying PEX cell cycle G0/G1 phase cell percentage rised to 81 .10%, S period cells decreased to 12 .04%, statistical disclosed significance （ P 〈0.05 ）.Conclusion PEX gene in C6 glioma, and its expression of the stem cells in vitro can directly suppress the BTSCs proliferation .

关 键 词：PEX基因 胶质瘤干细胞 真核表达载体 

分 类 号：R739.41[医药卫生—肿瘤]