siRNA干扰CD73表达抑制人乳腺癌细胞MB-MDA-231与细胞外基质的黏附  

Transfer of siRNA against CD73 suppresses human breast cancer cell line MB-MDA-231 adhesion to extracellular matrix

作　　者：支秀玲[1] 周婷婷[1] 王丽[1] 赵凤娣[1] 殷莲华[1] 周平[1] 

机构地区：[1]复旦大学上海医学院生理与病理生理学系,上海200032

出　　处：《中国病理生理杂志》2007年第2期217-221,共5页Chinese Journal of Pathophysiology

基　　金：国家"211工程"重点学科建设基金资助项目

摘　　要：目的:探讨CD73对乳腺癌细胞MB-MDA-231与细胞外基质黏附的影响及其机制。方法:①构建CD73 siRNA表达载体,脂质体介导转染MB-MDA-231细胞。②流式细胞仪检测瞬时转染效率,G418筛选稳定转染克隆。③半定量RT-PCR和W estern b lotting检测转染细胞CD73 mRNA及蛋白表达。④细胞黏附实验检测MB-MDA-231细胞CD73基因表达抑制后与细胞外基质黏附力的改变。结果:①CD73 siRNA表达载体转染MB-MDA-231可有效抑制CD73 mRNA表达(抑制效率=91.0%),P<0.01。②CD73 siRNA表达载体转染MB-MDA-231可有效抑制CD73蛋白表达(抑制效率=79.3%),P<0.01。③干扰MB-MDA-231细胞CD73表达可显著抑制细胞与细胞外基质的黏附(P<0.01)。结论:①CD73 siRNA表达载体可有效干扰MB-MDA-231细胞CD73基因表达。②MB-MDA-231细胞CD73基因表达抑制可显著降低细胞与细胞外基质的黏附。AIM： To study the effect of CD73 on breast cancer cell line MB - MDA -231 adhesion to extracellular matrix. METHODS：① CD73 siRNA plasmid was constructed and transfected into MB -MDA -231 cells by lipofectamine 2000. ② The transfection efficiency was analyzed by flow cytometry using eGFP as a marker gene. Stable transfected MB - MDA -231 cells were selected using G418. ③ RT - PCR and Western blotting analysis of CD73 expression in MB - MDA -231 cells was performed. ④The effects of CD73 on MB - MDA -231 cells adhesion to extracellular matrix were assessed by cell adhesion assay, RESULTS： ① Transfection of MB - MDA -231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at mRNA level. The efficiency was up to 91%. ② Transfection of MB -MDA -231 cells with CD73 siRNA plasmid significantly inhibited CD73 expression at a protein level. The efficiency was up to 79. 3%. ③ Treatment of MB -MDA -231 cells with CD73 siRNA resulted in diminished adhesion to extracellular matrix. CON- CLUSION = This study demonstrates that CD73 siRNA effectively inhibits CD73 gene expression in MB - MDA -231 cells leading to adhesion to extracellular matrix suppression.

关 键 词：5’-核苷酸酶 腺苷 RNA干扰 细胞粘附 

分 类 号：R730.23[医药卫生—肿瘤]