针对ERα的RNA干涉对人乳腺癌Bcap-37细胞生长的抑制作用  

Inhibitory effect of ERα-targeted RNA interference on the growth of human breast cancer cell line Bcap-37

作　　者：付海京[1] 贾林涛[1] 鲍炜[2] 赵晶[1] 孟艳玲[2] 王成济[1] 杨安钢[2] 

机构地区：[1]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 [2]第四军医大学基础部免疫学教研室,陕西西安710033

出　　处：《第四军医大学学报》2006年第15期1345-1348,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(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(30500274)

摘　　要：目的:探讨针对雌激素受体(ERα)分子的RNA干涉对肿瘤细胞生长的影响.方法:人工合成编码siRNA的脱氧寡核苷酸链,经磷酸化后退火连接进pSUPER载体.挑取和扩增序列正确的重组载体,稳定转染Bcap-37细胞后分别以RT-PCR和间接免疫荧光检测mRNA及蛋白质的表达情况,MTT法检测RNAi对细胞生长的影响.结果:siRNA表达载体转染后ERα表达水平明显降低,细胞增殖减慢.结论:针对ERα的RNA干涉可在体外抑制Bcap-37细胞生长.AIM： To investigate the inhibitory effect of ERα-targeted RNA interference （RNAi） on the growth of human breast cancer cell line Bcap-37. METHODS： ERa-targeted hairpin small interfering RNA （siRNA） genes, sierl and sier2, were obtained by oligonucleotide synthesis and annealing of the complementary single strand DNAs. PcDNA3 vector harboring each of the above genes was transfected into Bcap-37 cells in vitro. The expression of ERα gene in the transfected cells was examined by reverse transeriptase PCR （RT-PCR） and indirect immunofluorescence assay, and the effect of the targeted siRNA on cell growth and proliferation was evaluated by MTT assay. RESULTS： Both RT-PCR and indirect immunofluorescence assay revealed a remarkable decrease of ERα expression in pcDNA3- sierl-transfected cells, but not in pcDNA3 vector transfected cells. Cell proliferation was strongly inhibited after the expression of the ERa-targeted siRNAs as shown by microscopic observation and MTF. CONCLUSION： ERa-targeted RNAi in Bcap-37 cells can effectively inhibit the expression of ERα gene and the proliferation of the cells in vitro.

关 键 词：雌激素受体Α RNA干涉 基因治疗 

分 类 号：R730.5[医药卫生—肿瘤]