U251细胞系中脑肿瘤干细胞耐药性的初步研究  

Analysis of chemoresistance of brain tumor stem cell(BTSC) in U251 cell line

作　　者：邓永文[1] 周向阳 张明宇[3] 方加胜[3] 黄萌异[1] 舒毓高[1] 

机构地区：[1]湖南省人民医院神经外科,湖南长沙410005 [2]湖南省长沙市四医院神经外科,410078 [3]中南大学湘雅医院神经外科,湖南长沙410008

出　　处：《中国现代医学杂志》2011年第7期753-758,765,共7页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:30600636);湖南省科技厅科技计划一般项目(No:08fj3131);中南大学研究生教育创新工程博士研究生学位论文创新选题(No:76231)

摘　　要：目的探讨脑肿瘤干细胞对抗肿瘤药物的耐药性及其机制。方法通过磁珠分选(MACS)将U251细胞中的CD133+与CD133-细胞进行分选,用MTT法和流式细胞技术比较CD133+与CD133-细胞对抗肿瘤药物替尼泊苷(Vm-26)、卡莫司汀(BCNU)和顺铂(DDP)的敏感性、细胞凋亡率及细胞内药物蓄积的差异性,通过RT-PCR检测CD133+与CD133-细胞耐药基因谱的差异。结果 U251细胞系中,CD133+细胞约占5.4%,分选后的CD133+细胞的阳性率可达92.4%,CD133-细胞对抗肿瘤药物替尼泊苷、卡莫司汀和顺铂的敏感性明显高于CD133+细胞;流式细胞仪试验显示,在中等浓度的替尼泊苷作用下,CD133-细胞的凋亡明显,并出现明显的"亚G1峰",而CD133+细胞并未受到明显抑制和杀灭;流式细胞仪检测显示CD133+细胞内的药物蓄积明显低于CD133-细胞;RT-PCR实验显示MDR1、BCRP、MRP、MGMT和Bcl-2表达在CD133+明显高于CD133-细胞。结论 CD133+是U251细胞系中的耐药细胞亚群,高表达ABC转运体,DNA修复和抗凋亡基因可能是CD133+细胞耐药的重要机制。【Objective】 The purpose of this study was to investigate the drug sensitivity of BTSC.【Methods】 In this study,CD133＋and CD133-subpopulation cells in the human GBM U251 cell line were sorted by MACS.MTT and flow cytometry were performed to analyze the drug sensitivity of the two subpopulation to anti-tumor drug,Teniposide,Carmustine and Cisplatin.Finally,ABC transporter,DNA repair and apoptosis gene expression of the two subpopulation were analyzed by RT-PCR.【Results】 In this study,CD133＋cell occupied only 5.4% in the total population of U251 cell line,and the purity of CD133＋cells in the CD133＋population reached 92.4% after cell-sorting.Anti-tumor drug assay showed that,CD133-cells were more sensitive to Teniposide,Carmustine and Cisplatin than CD133＋cells.Treated with Teniposide,apoptotic percentage of CD133-cells was markedly higher than CD133＋cells,and ＂sub-G1peak＂ appeared in CD133-subpopulation while CD133＋did not,showed by FCM.And FCM also showed that,accumulation of anti-tumor drug in CD133＋cells was remarkably lower than CD133-cells.RT-PCR showed that,CD133＋ cells expressed remarkably higher than CD133-cells in MDR1,BCRP,MRP1,MGMT and Bcl-2.【Conclusions】 The subpopulation of CD133＋cells was the chemoresistant subpopulation in U251 cell line.High expression of ABC transporter,DNA repair and antiapoptosis may be the important chemoresistance mechanism of CD133＋cells.

关 键 词：肿瘤干细胞 磁珠分选 耐药性 U251人胶质母细胞瘤细胞系 

分 类 号：R739.4[医药卫生—肿瘤]