靶向VEGF小干扰RNA协同5-FU诱导人乳腺癌细胞MCF-7凋亡机制的研究  

Mechanism of the Apoptosis of Human Breast Cancer Cell Line MCF-7 Induced by Target-Oriented Vascular Endothelial Growth Factor siRNA Combined with 5-FU

作　　者：王誉静 王学祥[1,2] 于红[1] 王静[1] 张文卿[1] 

机构地区：[1]青岛大学医学院医学微生物学教研室,山东青岛266071 [2]苏州大学唐仲英血液学研究中心,江苏苏州215002

出　　处：《微生物学杂志》2015年第2期22-26,共5页Journal of Microbiology

基　　金：青岛市公共领域科技支撑计划项目(09-1-1-39-nsh)

摘　　要：探讨慢病毒介导的靶向VEGF小干扰RNA联合应用化疗药物5-FU诱导人乳腺癌细胞MCF-7凋亡的机制。以携带VEGF siRNA的慢病毒载体感染MCF-7细胞,应用RT-PCR和Western blot分别检测各组VEGF mRNA、VEGF蛋白及凋亡相关蛋白的表达,流式细胞术检测细胞凋亡。结果表明,慢病毒VEGF siRNA干扰组细胞VEGF mRNA和蛋白表达水平明显低于对照组,凋亡相关蛋白P53及P21表达上调,而SIRT1、Bcl-2及Survivin表达下调。流式细胞术检测显示慢病毒干扰组及5-FU组细胞凋亡率显著升高,联合治疗组的协同作用更为明显。上述结果表明:慢病毒介导的RNA干扰能明显抑制MCF-7细胞VEGF的表达,通过下调SIRTI蛋白的表达,导致P53蛋白表达上调,并调控其下游P21、bcl-2和Survivin的表达,从而诱导MCF-7细胞的凋亡,并且提高了MCF-7对5-FU的敏感性。The mechanism of the apoptosis of human breast cancer cell line MCF-7 induced by lentivirus with media- tion of target-oriented VEGF siRNA combined with the anticancer drug 5-FU was investigated. Lentiviral vectors carry- ing VEGF siRNA was infected into human breast cancer MCF-7 cells, the level of VEGF mRNA, VEGF protein and apoptosis related proteins of each group were measured by RT-PCR and Western blot respectively, cell apoptosis was analyzed by flow eytometry. The results showed that in the lentivirus VEGF siRNA interference group the VEGF mR- NA and protein expression level was significantly lower than that in the control group, the expression of the apoptosis related proteins P53 and 1〉21 increased, while SIRT1, Bcl-2 and Survivin decreased. Flow cytometry results showed that the cell apoptosis rate in the interference group and 5-FU group were increased significantly, the synergistic effect of the combined treatment group was more obvious. The above-mentioned results indicated that lentivirus mediated siRNA interfering could significantly suppressed VEGF expression in MCF-7 cell, induced the apoptosis of MCF-7 cells by down-regulating the expression of SIRT1, and induced up-regulating the expression of P53, which regulated the expression of its down stream P21, Bcl-2 and Survivin, and induced the apoptosis of MCF-7, moreover improved the sensitivity of MCF-7 to 5-FU.

关 键 词：VEGF RNA干扰 乳腺癌细胞 5-FU 细胞凋亡 

分 类 号：Q939.91[生物学—微生物学] R737.9[医药卫生—肿瘤]