RNAi靶向沉默乳腺癌细胞系MCF-7中EphA2表达的实验研究

目的探讨逆转录病毒介导的RNAi对乳腺癌细胞系MCF-7中EphA2表达的影响。方法构建重组的表达EphA2siRNA的逆转录病毒载体,将重组载体和空载体转染入MCF-7细胞中,筛选出稳定转染的单克隆细胞扩大培养,并用Westen blot检测EphA2蛋白的表达情况。结果 Western blot分析,与对照空载体及未转染细胞比较,重组逆转录病毒载体转染的MCF-7细胞中EphA2蛋白表达水平明显降低(P<0.05)。结论通过逆转录病毒载体介导的RNAi,能够抑制MCF-7细胞中EphA2的蛋白表达,为观察转染siRNA的逆转录病毒表达载体的乳腺癌细胞恶性生物学行为变化奠定了实验基础,为以EphA2为靶向的乳腺癌的基因治疗提供了新的思路和手段。

Study of targeted silencing of EphA2 in breast cancer cell line MCF-7 by RNA interference

Objective To investigate the effects of retrovirus-mediated siRNA targeting EphA2 on the breast cancer cell line MCF-7.Methods The recombinant retroviral vector expressing EphA2 short interference RNA（siRNA） was constructed and transfected into the MCF-7 cells,and then stable transfectants were isolated by puromycin and were cultured.The EphA2 protein in transfected MCF-7 cells was detected by Western blot.Results Compared with cells transfected with empty retroviral vector and untransfected cells,the level of EphA2 protein expression was greatly decreased in MCF-7 cells transfected by the recombinant retroviral vector with Western blot techniques.Conclusions EphA2 protein expression in MCF-7 cell line is successfully suppressed by using the recombinant retrovirus-mediated RNAi technique,indicating that this may contributes to the study of the changes of malignant biological activity of breast cancer cell lines by transfection of siRNA retrviral vectors and provide a new solution for EphA2-targeting therapeutic intervention of breast carcinoma.