ABCE1基因沉默对人乳腺癌细胞MCF-7增殖、凋亡的影响  

Effects of ABCE1 Gene Silence in Proliferation and Apoptosis of Human Breast Cancer Cells MCF-7

作　　者：龚向南[1] 黄波[1] 周红丽[2] 

机构地区：[1]辽宁医学院附属第一医院胸外科,锦州121000 [2]辽宁医学院附属第一医院肾内科,锦州121000

出　　处：《中国细胞生物学学报》2013年第9期1308-1313,共6页Chinese Journal of Cell Biology

基　　金：辽宁省科技厅自然科学基金(批准号:201202143);辽宁医学院博士启动基金(批准号:Y2011B04)资助的课题~~

摘　　要：为研究ABCE1基因沉默对人乳腺癌MCF-7细胞增殖、凋亡的影响并初步探讨其机制,将构建好的ABCE1的siRNA绿色荧光载体转染入乳腺癌细胞MCF-7中,于荧光显微镜下观测转染效率,运用Western blot及RT-PCR实验证实基因沉默的效果,同时应用Western blot检测RNase L蛋白的表达,MTT法分析细胞增殖能力并绘制细胞生长曲线,流式细胞术检测细胞凋亡率。结果显示,将构建好的载体成功转入了乳腺癌细胞MCF-7。转染ABCE1-siRNA后细胞活性降低,ABCE1的表达受到阻断(P<0.01),RNase L蛋白含量明显增加(P<0.01),细胞的增殖能力明显降低而凋亡率显著增加(P<0.01)。证实:ABCE1-siRNA可成功沉默乳腺癌MCF-7细胞ABCE1基因的表达,沉默该基因可抑制人乳腺癌细胞MCF-7增殖并诱导细胞凋亡,其机制可能与其阻断了2-5A/RNase L细胞通路有关。This paper aim at investigating the effects of ABCE1 gene silence in proliferation and apoptosis of human breast cancer cells MCF-7 and discuss its mechanism preliminarily. The siRNA vector of constructed ABCE1 was transfected into MCF-7. The efficiency of transfection was observed under the fluorescence microscope. Effects of gene silencing was confirmed by Western blot and RT-PCR. Expressions of RNase L protein was examined by Western blot. The proliferation was analysised by MTT and growth curve was drew. The apoptosis was analysised by flow cytometry. Results showed that ABCEI-siRNA was transfected into MCF-7 successfully. After transfection, cell viability reduced and expression of ABCE1 were blocked （P〈0.01）. The expression of RNase L increased obviously （P〈0.01）. The ability of cell proliferation decreased but cell apoptosis rate increased （P〈0.01）. From discussion we can draw the conclusion that ABCEI-siRNA can successfully silence ABCE1 gene in human breast cancer cells MCF-7. Silencing this gene can restrain the proliferation of human breast cancer cells MCF-7 and induce apoptosis. The mechanism may be related to blocking-up cell pathway of 2-5A/RNase L.

关 键 词：乳腺癌 MCF-7 小干扰RNA ABCE1 核糖核酸酶L 

分 类 号：R737.9[医药卫生—肿瘤]