RNA干扰技术抑制乳腺癌细胞系MDA-MB-231中SATB1的表达  

RNAi-Mediated Gene Silencing of SATB1 Expression in MDA-MB-231 Breast Cancer Cell Line

作　　者：张瑞[1] 曹以诚[1] 张珍武[1] 杜淑敏[1] 徐爱群[1] 

机构地区：[1]华南理工大学生物科学与工程学院,广东广州510006

出　　处：《现代食品科技》2010年第7期669-672,687,共5页Modern Food Science and Technology

基　　金：国家自然科学基金重大研究计划(编号:90412015);教育部中国网格计划生物信息网格平台子项目(编号:B1-137040130)

摘　　要：构建含有靶向satb1基因的siRNA质粒,体外观察对乳腺癌细胞系MDA-MB-231的SATB1mRNA以及蛋白质表达的影响。设计合成三对靶向人源satb1基因的siRNA,并分别克隆在质粒载体上,在脂质体的介导下转染高表达SATB1的乳腺癌细胞系MDA-MB-231。运用RealTime-PCR方法分析SATB1mRNA的表达情况,Western-Blot方法检测蛋白的表达量。结果表明:经过酶切鉴定和测序证实三个重组质粒psiS823、psiS2567、psiS3373分别构建成功,转染乳腺癌细胞系MDA-MB-231能明显抑制SATB1mRNA及蛋白的表达。说明本试验成功构建的重组质粒psiS823、psiS2567、psiS3373都能有效降低的人乳腺癌细胞MDA-MB-231中SATB1的表达,为SATB1高表达的乳腺癌基因治疗提供了新方法。An expression vector of siRNA of human satb1 was constructed and its effects on SATB1 expression in human breast cancer cell line MDA-MB-231 in vitro were investigated in this paper. Three pairs of siRNA targeting human satb1 were designed, synthesized and cloned in the plasmid vector. Then these recombinant plasmids were transfected by liposome into highly SATB1-expressing MDA-MB-231 cells. Real Time-PCR and Western-blot method were used to analyze SATB1 mRNA expression and its protein product, respectively. The analysis of enzyme digestion and sequencing demonstrated that the recombinant plasmidhas psiS823, psiS2567, psiS3373 had been constructed successfully. And their transfection into human breast cancer cells MDA-MB-231 can significantly reduce the expression of SATB1 mRNA and it’s protein product. It was conclude that recombinant plasmid psiS823, psiS2567, psiS3373 can significantly reduce SATB1 expression of human breast cancer cell MDA-MB-231. This research provided a new strategy for gene therapy for breast cancers with high SATB1 expression.

关 键 词：乳腺癌 SABT1 SIRNA 基因表达 

分 类 号：R737.9[医药卫生—肿瘤]