KDR靶向RNA干扰对MCF-7细胞凋亡的影响(英文)

Apoptosis Effect of RNAi Targeting KDR on MCF-7 Cells

目的:研究KDR靶向RNA干扰对MCF-7细胞凋亡的影响,探讨其可能的机制。方法:采用阳离子脂质体Lipofecta-mine2000TM作为转染试剂将人KDR基因的siRNA转染人类乳腺细胞株MCF-7,诱RNAi,采用Hoechst33258染色和半定量RT-PCR检测Caspase-3、survivin的mRNA表达及细胞凋亡变化;比色法检测Caspase-3的活性;利用免疫组织化学方法检测survivin的表达,并用图像分析仪分析蛋白表达强度。结果:靶向KDR的siRNA转染MCF-7后,Caspase-3的mRNA表达上调,survivin基因mRNA及蛋白表达水平下调(P<0.05)。结论:KDRsiRNA通过减少乳腺癌细胞survivin的表达,增加Caspase-3表达来促进肿瘤细胞凋亡,发挥其抗肿瘤作用。

Objective： To investigate the apoptosis effect of small interfering RNA （siRNA） targeting kinase insert domain-containing receptor （KDR） on MCF-7 cells and to investigate the possible mechanisms. Methods： KDR-siRNA was chemically synthesized and transfected into MCF-7 cells by cationic liposome Lipofectamine2000TM. Cell apoptosis was measured by Hoechst33258 staining; The expression ofCaspase-3 mRNA and survivin mRNA was detected by semi-quantitative RT-PCR; Caspase-3 activity was measured by colorimetry; The expression of survivin protein was detected by immunohistochemistry and analyzed by the image analyzer. Result.s： siRNA effectively knocked-down KDR gene. KDR-siRNA enhanced the expression of Caspase-3 and down-regulated the expression of survivin （p〈0.05）. Conclusions： KDR-siRNA induced tumor cells apoptosis by increasing the expression of Caspase-3 and decreasing the expression of survivin.