BSP基因RNA干扰对乳腺癌MDA-MB-231BO细胞生物学特性的影响  

Effects of BSP RNAi on Human Breast Cancer Cell Line MDA-MB231BO

作　　者：燕慧[1,2] 王捷[2] 杨自飞[2] 张宏斌[2] 夏冰[2] 冼江[2] 王江涛[2] 

机构地区：[1]华南理工大学生物科学与工程学院,广州510006 [2]广州军区广州总医院医学实验科,广州510010

出　　处：《生物技术通报》2010年第6期142-145,共4页Biotechnology Bulletin

基　　金：广东省自然科学基金重点项目(06104396)

摘　　要：旨在研究RNA干扰(RNA interference,RNAi)抑制骨唾液酸蛋白(bone sialoprotein,BSP)基因表达后对人乳腺癌细胞MDA-MB-231BO生物学特性的影响。应用pSilencer5.1-U6Retro构建针对BSP基因的siRNA逆转录病毒重组表达质粒,将重组质粒转染293细胞制备病毒悬液感染MDA-MB-231BO细胞,利用嘌呤霉素筛选抑制BSP表达的乳腺癌细胞。Western blotting检测细胞内BSP蛋白表达,采用MTT法和集落形成试验检测细胞的增殖,流式细胞仪检测细胞周期变化。结果显示,成功构建BSP基因RNAi稳定转染的231BO-BSP27细胞。231BO-BSP27细胞内BSP蛋白表达抑制率为69.3%,与对照组细胞相比,231BO-BSP27细胞的生长速率和克隆形成率明显降低;S期细胞数量明显减少,G0/G1期细胞增多。由此证实,逆转录病毒介导的RNAi能实现BSP基因稳定沉默,从而抑制MDA-MB-231BO细胞的生长和增殖。The object was to study the effects of BSP RNAi on the biologic characteristics of human breast cancer line MDA-MB-231BO.BSP RNAi retrovirus vector pSilencer-BSP27 was constructed and packaged into 293 cells,and the MDA-MB-231BO cells were infected with the retrovirus.Finally,the cells inhibited BSP expression were screened by puromycin.The expression of BSP was analyzed by Western blotting.Cell proliferation was detected by MTT assay and colony formation assay,the cell cycle phases were investigated by flow cytometry.Results indicated that pSilencer-BSP27 stably transfected MDA-MB-231BO cells（231BO-BSP27）was successfully constructed.Expression of BSP was reduced 69.3%in 231BO-BSP27 cells.The growth rate and colony formation ability were both significantly lower in 231BO-BSP27 cells than those in the mock and negative control group.Furthermore,the cell cycle of 231BO-BSP27 arrest at G0/G1 phase with a significant decrease of cells in S phase.Therefore,it proved that Retrovirus-mediated BSP RNAi can result in stably silencing of BSP gene,inhibit the growth and proliferation in MDA-MB-231BO cells.

关 键 词：骨唾液酸蛋白 RNA干扰 增殖 细胞周期 

分 类 号：R737.9[医药卫生—肿瘤]