shRNA抑制C-erbB-2基因表达对小鼠Lewis细胞凋亡的影响  

Effect of C-erbB-2 gene inhibition by shRNA on apoptosis of Lewis cells

作　　者：曹新梅[1] 张岱权[2] 夏纪毅[3] 王栩[1] 

机构地区：[1]泸州医学院免疫学教研室,四川泸州646000 [2]泸州医学院附属医院中医科,四川泸州646000 [3]泸州医学院药物与功能性食品研究中心,四川泸州646000

出　　处：《泸州医学院学报》2014年第4期345-348,共4页Journal of Luzhou Medical College

基　　金：四川省卫生厅基金(100236)

摘　　要：目的:探讨shRNA抑制C-erbB-2基因表达对小鼠肺腺癌Lewis细胞凋亡的影响。方法:构建4个pGPU6/RFP/Neo-erbB-2质粒和pGPU6/RFP/Neo-shNC。应用Lipofectamine 2000转染pGPU6/RFP/Neo-shNC至小鼠肺腺癌Lewis细胞,应用荧光显微镜观察荧光,确定最佳转染效率。应用RT-PCR检测C-erbB-2 mRNA的水平,应用Western blotting检测该蛋白的表达,筛选干扰效果最好的质粒。应用流式细胞术检测空白组、pGPU6/RFP/Neo-shNC组和最佳干扰组细胞的凋亡率。结果:pGPU6/RFP/Neo-shNC(μg):Lipofectamine(μl)为0.4:1时,转染效果最好。4个pGPU6/RFP/Neo-erbB-2质粒均能降低C-erbB-2 mRNA水平和蛋白的表达,其中干扰效果最好的是pGPU6/RFP/Neo-erbB-2-mus-3669,用于后续实验。pGPU6/RFP/Neo-erbB-2-mus-3669质粒组的凋亡率高于空白组和pGPU6/RFP/Neo-shNC组(P<0.05)。结论:pGPU6/RFP/NeoerbB-2-mus-3669质粒能抑制小鼠Lewis细胞C-erbB-2基因的表达,诱导细胞凋亡。Objective： To investigate the effect of C-erbB-2 gene inhibition by shRNA on the apoptosis of Lewis cells. Methods： 4 pGPU6/RFP/Neo-erbB-2 plasmid and pGPU6/RFP/Neo-shNC were synthesized.pGPU6/RFP/Neo-shNC was transfected into mouse Lewis cells to test the transfection efficiency. The interf-erence effect was tested by RT-PCR and Western blotting to select the best effective C-erbB-2 shRNA. The apoptotic rate of cells was tested by FCM. Results： The transfection efficiency was the best when pGPU6/RFP/Neo-shNC（μg）： Lipofectamine（μl） was 0.4：1. pGPU6/RFP/Neo-erbB-2-mus-3669 had the best interference effect, and it could induce higher apoptotic rate of Lewis cells than control group and pGPU6/RFP/Neo-shNC group（P 0.05）. Conclusion： pGPU6/RFP/Neo-erbB-2-mus-3669 can inhibit the expression of C-erbB-2 gene and induce the apoptosis of Lewis cells.

关 键 词：C-ERBB-2 SHRNA Lewis细胞 凋亡 

分 类 号：R734.2[医药卫生—肿瘤]