PRL-3基因沉默对乳腺癌MCF-7细胞增殖和凋亡的影响  

Effects of PRL-3 gene on the proliferation and apoptosis of breast cancer MCF-7 cells

作　　者：钟琰[1] 吴爱国 纪术峰[1] 沈三弟[1] 

机构地区：[1]南方医科大学珠江医院普外科,510282

出　　处：《新医学》2011年第6期370-374,共5页Journal of New Medicine

基　　金：广东省科技计划资助项目(2008B030301345)

摘　　要：目的:观察短发夹状小干扰RNA(shRNA)沉默PRL-3基因对人乳腺癌MCF-7细胞增殖、凋亡的影响。方法:构建PRL-3基因特异性shRNA表达载体,使用脂质体法将PRL-3-shRNA表达载体转染入MCF-7细胞。采用实时荧光定量PCR和蛋白免疫印迹法检测转染后MCF-7细胞PRL-3基因mRNA和蛋白的表达;运用MTT法检测转染后MCF-7细胞的增殖水平,流式细胞仪检测细胞凋亡情况。结果:酶切鉴定和测序分析证实PRL-3-shRNA表达载体成功构建。转染成功后,PRL-3-shRNA组PRL-3基因mRNA和蛋白表达水平明显降低。MTT结果显示MCF-7细胞转染PRL-3-shRNA后细胞增殖水平降低;流式细胞仪检测显示,PRL-3-shRNA转染后,MCF-7细胞凋亡率明显增高。结论:沉默PRL-3基因表达可抑制MCF-7细胞的增殖,促进其凋亡。Objective：To study the effects of short hairpin RNA（shRNA） targeting PRL-3 gene on the proliferation and apoptosis of breast cancer MCF-7 cells.Methods：The PRL-3 specific shRNA expression vector was constructed and confirmed by sequencing analysis.PRL-3-shRNA expression vector was transfected into MCF-7 cells via lipofectamineTM 2000,and the expression levels of PRL-3 mRNA and protein compared with those non-transfected cells were determined by real-time fluorescent quantitative PCR and western blotting.Flow cytometry and MTT assay were performed to assess the effects of PRL-3-shRNA on the proliferation and apoptosis of MCF-7 cells,respectively.Results：PRL-3-shRNA expression vector was confirmed correct by restriction enzyme digestion and sequencing.After transfection,PRL-3 mRNA and protein expression levels were decreased significantly.MTT results showed that the proliferation of MCF-7 cells was markedly suppressed,and flow cytometry results indicated that the apoptotic rate of MCF-7 cells was significantly increased.Conclusion：Silencing the expression of PRL-3 gene by shRNA not only inhibits the proliferation of MCF-7 cells,but also promotes its apoptosis.

关 键 词：乳腺癌 PRL-3 SHRNA 基因治疗 细胞增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]