HER2基因沉默抑制人胃癌细胞SGC-7901的生长和侵袭  

HER2-targeted RNA interference inhibits growth and invasion of human gastric carcinoma cell SGC-7901

作　　者：鲍炜[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] 杨安钢[1] 

机构地区：[1]第四军医大学基础医学部免疫学教研室 [2]第四军医大学基础医学部生物化学与分子生物学教研室 [3]第四军医大学附属西京医院呼吸内科,西安710032

出　　处：《中国肿瘤生物治疗杂志》2007年第4期328-332,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家重点基础研究发展(973)计划资助项目(No2004CB518805);国家自然科学基金资助项目(No30500592);教育部"长江学者和创新团队发展计划"资助项目(IRT0459)~~

摘　　要：目的:探讨针对HER2基因的RNA干涉对人胃癌细胞生长和侵袭的影响。方法:构建了针对癌基因HER2的siRNA表达载体,分别命名为pcDNA3-sihe1和pcDNA3-sihe2,将构建的干涉载体与对照载体转染胃癌SGC-7901细胞,通过间接免疫荧光实验确定干涉效果;以软琼脂集落形成实验检测干涉后细胞的集落形成能力,以流式细胞术检测细胞在悬浮培养条件下的凋亡情况;并以Boyden chamber法检测转染后细胞的侵袭能力。结果:成功构建针对癌基因HER2的siRNA表达载体,siRNA表达载体转染SGC-7901细胞后HER2表达水平明显降低,证实siRNA成功抑制HER2蛋白的表达。HER2低表达的细胞株集落形成能力降低,流式细胞术检测发现HER2下调可引起胃癌细胞的凋亡,Boydenchamber实验表明针对HER2的RNA干涉可抑制胃癌细胞SGC-7901的侵袭能力。结论:RNA干涉有效地抑制了HER2分子的表达,进而影响SGC-7901细胞的生长和侵袭,本实验为进一步研究HER2分子与肿瘤细胞生长和转移的关系奠定了基础。Objective： To investigate the effect of HER2-targeted RNA interference （RNAi） on growth and invasion of human gastric cancer cell line SGC-7901. Methods： HER2-targeted hairpin small interfering RNA （siRNA） genes, pcD- NA3-sihel and pcDNA3-sihe2, were obtained by oligonucleotide synthesis and annealing of the complementary single strand DNAs; they were used to transfect SGC-7901 cells in vitro. The expression of HER2 gene in the transfected cells was examined by indirect immunofluorescence assay and the effects of the HER2-targeted siRNA on cell growth, apoptosis and invasion were evaluated by colony formation assay, flow cytometry, and Boyden chamber assay. Results： We success- fully constructed siRNA expression vector targeting HER2 gene. Indirect immunofluorescence assay revealed a remarkable decrease of HER2 expression in pcDNA3-sihel -and pcDNA3-sihe2-transfected cells, but not in pcDNA3 vector transfected cells. The colony-forming ability of cells with low expression of HER2 was decreased. Flow cytometry demonstrated downregulation of HER2, with resulted in apoptosis of gastric cancer cells. Boyden chamber assays revealed that HER2-targeted RNAi suppressed the invasion ability of SGC-7901 cells. Conclusion： HER2-targeted RNAi can effectively inhibit the expression of HER2 in SGC-7901 cells and subsequently influence the growth and invasion of SGC-7901 ceils, which paves a way for studying the relationship between HER2 molecular and tumor growth and metastasis.

关 键 词：HER2基因 RNA干涉 胃肿瘤 侵袭 

分 类 号：R730.5[医药卫生—肿瘤]