HER2 siRNA抑制肺癌细胞增值的实验研究  

Inhibition of cell proliferation of lung cancer by vector based-small interfering RNA targeting HER2 gene

作　　者：任新玲[1] 钱桂生[1] 付海京[2] 鲍炜[3] 杨安钢[4] 

机构地区：[1]第三军医大学新桥医院全军呼吸内科研究所,重庆400037 [2]第四军医大学西京医院呼吸内科 [3]第四军医大学基础部生物化学与分子生物学教研室 [4]第四军医大学基础部免疫学教研室

出　　处：《中华肺部疾病杂志（电子版）》2007年第1期16-18,共3页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：国家重大基础研究计划项目"973"(2004CB518805);教育部"长江学者和创新团队发展计划"项目(IRT0459)

摘　　要：目的构建针对HER2的RNA干涉载体,观察对HER2表达抑制及抑制后对非小细胞肺癌生长的影响。方法设计和合成长度为64nt的脱氧寡核苷酸链,退火互补成双链,克隆至pcD-NA3质粒载体,转化DH5α菌株,提取质粒,转染过表达HER2的肺腺癌细胞系SPC-A-1,RT-PCR检测HER2的mRNA表达,Western Blot和间接免疫荧光法检测HER2的蛋白表达,FCM分析细胞周期变化,MTT法观察细胞生长。结果成功构建了HER2特异性RNA干涉载体pcDNA3～siHER2;瞬时转染SPC-A-1细胞,HER2的mRNA及蛋白表达均降低;G_1期细胞较亲代增加9.9%;肿瘤细胞增殖速度减慢(P<0.05)。结论成功构建了HER2 RNA干涉载体,转染后可有效降低肺腺癌细胞系SPC-A-1 HER2的mRNA转录及蛋白水平表达,阻滞转染细胞于G_1期,造成转染细胞生长速度减慢,这有望为肺癌基因治疗提供一种选择手段。Objective To construct RNA interference vector to down-regulate HER2 gene and to study the RNA interference effect on the cell cycle and tumor growth of non-small-cell lung cancer （NSCLC）. Methods Oligonucleotides of 64 base pairs for hairpin RNA targeting HER2, designed, chemically synthesized, and annealed, were cloned into pcDNA3 vector. After identification by restriction digestion, the right vectors were transiently transfected into SPC-A-1 cell, a human lung adenocarcinoma cell line. HER2 mRNA and HER2 protein were detected by RT-PCR, and Western Blot, respectively. FCM analysis and MTT method were applied to measure cell cycle and cell growth. Results The vector of RNA interference which could interfere HER2 gene was successfully constructed, which could increase the number of cells in G0/G1 phase by 9.9% and inhibit the tumor cell growth. Conclusion We have successfully constructed expressing hairpin RNA against HER2 vector. The vector can effectively silence HER2 gene, increase the number of cells in G0/G1 phase, and then slow down the growth of tumor cells. This may be a useful therapeutic strategy for HER2 over-expressing NSCLC.

关 键 词：SIRNA RNA 干涉 非小细胞肺癌 HER2 

分 类 号：R734.2[医药卫生—肿瘤] R-33[医药卫生—临床医学]