靶向于HER2的siRNA表达载体在人乳腺癌细胞SKBr-3中的表达  

Expression of siRNA vector against HER2 in the human breast carcinoma SKBr-3 cells

作　　者：鲍炜[1] 张立红[1] 付海京[2] 贾林涛[2] 张勇[1] 刘家云[1] 任新玲[3] 温伟红[1] 孟艳玲[1] 王成济[2] 杨安钢[1,4] 

机构地区：[1]第四军医大学基础部免疫学教研室 [2]第四军医大学基础部生物化学与分子生物学教研室 [3]第四军医大学西京医院呼吸内科 [4]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710033

出　　处：《第四军医大学学报》2007年第15期1355-1358,共4页Journal of the Fourth Military Medical University

基　　金：国家重点基础研究发展(973)计划项目(2004CB518805);教育部"长江学者和创新团队发展计划"(IRT0459);国家自然科学基金(30500592)

摘　　要：目的:探讨针对HER2分子的RNA干涉对乳腺癌细胞生长的影响.方法:人工合成DNA,经退火磷酸化后克隆入pSUPER载体.基因转染后通过观察细胞形态和细胞计数等方法验证发夹状小干涉RNA(siRNA)对细胞生长状态的影响,以激光共聚焦检测基因表达情况.结果:siRNA表达载体转染后可以引起SKBr-3细胞大量死亡,间接免疫荧光检测出HER2蛋白表达降低.结论:针对HER2进行RNA干涉可在体外抑制SKBr-3细胞生长.AIM： To investigate the effect of HER2-targeted RNA interference （RNAi） on the growth of human breast carcino- ma SKBr-3 cells. METHODS： HER2-targeted hairpin small in- terfering RNA （siRNA） genes were obtained by oligonucleotide synthesis and annealing of the complementary single strand DNAs. pSUPER vector harboring each of the above genes was transfected into SKBr-3 cells in vitro. The expression of HER2 gene in the transfected cells was examined by RT-PCR and indi- rect immunofluorescence assay, and the effect of the HER2-targe- ted siRNA on cell growth and proliferation was evaluated by cell counting and morphological observation. RESULTS： Both RT- PCR and indirect immunofluorescence assay revealed a remarkable decrease of HER2. expression in pSUPER-sihel- and pSUPER-si- he2-transfected cells, but not in pSUPER vector transfected cells. Cell proliferation was dramatically inhibited after the expression of the HER2-targeted siRNAs as shown by microscopic observation and cell counting. CONCLUSION： HER2-targeted RNAi in SK- Br-3 cells can effectively inhibit the expression of HER2 gene and the proliferation of the cells in vitro.

关 键 词：HER2 RNA干涉 基因治疗 

分 类 号：R730.5[医药卫生—肿瘤]