【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。更多还原

【Abstract】 The mechanism of the apoptosis of human breast cancer cell line MCF-7 induced by lentivirus with mediation of target-oriented VEGF siRNA combined with the anticancer drug 5-FU was investigated.Lentiviral vectors carrying 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 cytometry.The results showed that in the lentivirus VEGF siRNA interference group the VEGF mRNA and protein expression level was significantly lower than that in the control group,the expression of the apoptosis related proteins P53 and P21 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-7cells by down-regulating the expression of SIRTl,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.更多还原