4 669-672 转687 基础研究 4+20 RNA干扰技术抑制乳腺癌细胞系MDA-MB-231中SATB1的表达.pdf

现代食品科技 Modern Food Science and Technology 2010, Vol.26, No.7 RNA干扰技术抑制乳腺癌细胞系 MDA-MB-231中SATB1的表达 张瑞，曹以诚，张珍武，杜淑敏，徐爱群 （华南理工大学生物科学与工程学院，广东广州 510006） 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SATB1 mRNA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Real Time-PCR方法分析SATB1 mRNA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SATB1 mRNA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。 关键词：乳腺癌；SABT1；siRNA；基因表达 文章篇号：1673-9078(2010)7-669-672 RNAi-Mediated Gene Silencing of SATB1 Expression in MDA-MB-231 Breast Cancer Cell Line ZHANG Rui, CAO Yi-cheng, ZHANG Zhen-wu, DU Shu-ming, XU Ai-qun (School of Bioscience and Bioengineering South China University of Technology, Guangzhou 510006, China) Abstract: 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 conc