胶质瘤U251细胞及其干细胞抗凋亡与多重耐药基因的关系

目的利用培养成功的U251胶质瘤干细胞,探讨其抗凋亡和耐药基因的表达差异。方法WST-8法检测胶质瘤干细胞增殖活性,实时荧光定量RT-PCR技术检测Livin、Livinα、Livinβ、Survivin和MRP1、MRP3 mRNA的表达。结果单层培养的U251细胞比胶质瘤干细胞表现出更强的增殖活性;U251胶质瘤干细胞Livin、Livinα、Livinβ、Survivin和MRP-1 mRNA表达量较U251单层培养细胞表达量都有不同程度的升高,而MRP-3 mRNA表达量降低。结论U251胶质瘤干细胞抗凋亡和MRP-1 mRNA表达量较U251单层培养细胞表达量有不同程度的升高,提示胶质瘤干细胞比较其同源的肿瘤细胞具有更强的耐药性,这可能是肿瘤耐药的机制。

Expression of anti-apoptotic and multidrug resistance-associated protein genes in cancer stem cell isolated from U251 glioma cell line

Objective To isolate and culture cancer stem cells from U251 glioma cell line and detect the expression of anti-apoptotic and multidrug resistance-associated protein（MRP） genes.Methods Expression of livin,livinα,Livinβ,survivin,MRP1 and MRP3 on cancer stem cell isolated from U251 glioma cell line was assayed by real-time quantitative RT-PCR.Results The cancer stem cell derived from U251 glioma cell line was isolated successfully.These glioma stem cells maintained a sphere-like growth status in serum-free medium in vitro,and self-renew,proliferate.The mRNA expression of U251 cancer stem cells on livin,livinα,survivin,MRP1 was significantly up-regulated than that of U251 cells.However,the mRNA expression of MRP3 was down-regulated.Conclusion Cancer stem cells can be isolated from U251 glioma cell line.However,it is observed that the generating rate of tumor stem cells are lower than that of U251 cells,and the expression levels of anti-apoptotic and MRP1 genes are higher than that of the U251 cells,which show cancer stem cells are the factor to maintain tumor growth and resist apoptosis and to pump the anti-tumor drugs out of cells,and this is what we need further research on.