短发夹RNA沉默S100A4基因对乳腺癌MCF-7细胞体外增殖和迁移力的抑制  

Effects of Short Harpin in RNA Targeting S100A4 Gene on Proliferation and Migration Potential of MCF-7 Cells in vitro

作　　者：马雷[1] 吴爱国 纪术峰[1] 杨华峰[1] 

机构地区：[1]南方医科大学珠江医院普外科,广州510282

出　　处：《肿瘤防治研究》2010年第4期402-406,共5页Cancer Research on Prevention and Treatment

基　　金：广东省科技计划资助项目(2008B030301345)

摘　　要：目的观察靶向S100A4的shRNA对MCF-7细胞增殖和迁移力的影响。方法构建S100A4基因特异性shRNA表达载体,使用QRT-PCR和Western blot检测转染后MCF-7细胞中S100A4的表达水平,运用MTT法和流式细胞术检测转染后MCF-7细胞增殖水平和凋亡率的变化。将S100A4-shRNA表达载体转染入MCF-7细胞,G418筛选及克隆化培养,并运用划痕实验检测其迁移力的变化。结果经测序鉴定证实成功构建S100A4-shRNA表达载体。转染48h后,所构建的S100A4-shRNA表达载体能够有效抑制MCF-7细胞中S100A4的表达,并且可以显著降低MCF-7细胞增殖水平以及诱导细胞进入晚期凋亡;稳定转染S100A4-shRNA表达载体的MCF-7细胞形态无显著变化,但是迁移力明显下降。结论S100A4-shRNA表达载体能有效地抑制乳腺癌MCF-7细胞中S100A4的表达,从而降低细胞增殖水平和迁移力。Objective To study the effects of short hairpin in RNA（shRNA） targeting S100A4 on the proliferation and migration potential of breast cancer MCF-7 cells.Methods The S100A4-shRNA expression vector was constructed and confirmed by sequencing. Then the plasmid was transfected into MCF-7 cells via lipofectamine TM 2000, and the changes of S100A4 expression were determined using quantitative RT-PCR and Western blot 48 h after transfection. Flow cytometry and MTT assay were performed to assess the effect of the S100A4-shRNA expression vector followed by G418 selection,colon culture.Migration capability of stably transfected MCF-7 cells in vitro was evaluated by using wound assay.Results S100A4-shRNA expression vector was constructed and transfected into MCF-7 cells.48h after transfection, S100A4 mRNA and S100A4 protein level were decreased significantly. The suppression of S100A4 expression by S100A4-shRNA expression vector inhibited the growth of MCF-7 cells and significantly higher apoptosis cell rate was observed in S100A4shRNA expression vector-transfected cells.The morphology of stably transfected MCF-7 cells didn’t change ,but the potentiality of migration of MCF-7 cells was decreased significantly.Conclusion S100A4-shRNA expression vector can significantly suppress S100A4 expression and inhibit the proliferation while decreasing the migration capability of MCF-7 cells.

关 键 词：乳腺癌 S100A4 shRNA 增殖 迁移力 

分 类 号：R737.9[医药卫生—肿瘤]