针对HER2-neu的siRNA对非小细胞肺癌细胞生长的抑制作用.doc

针对HER2-neu的siRNA对非小细胞肺癌细胞生长的抑制作用.doc针对HER2/neu的siRNA对非小细胞肺癌细胞生长的抑制作用
作者:任新玲,钱桂生,鲍炜,付海京,贾林涛,张瑞,王涛,许彦鸣,杨安钢
【关键词】非小细胞肺
Inhibition of tumor groall cell lung cancer by small interfering RNA targeting to HER2/neu gene
【Abstract】 AIM: To construct RNA interference (RNAi) vector to door groall cell lung cancer. METHODS: Oligonucleotides of 64 base pairs for hairpin RNA targeting HER2/neu ically synthesized and annealed, and cloned into pSUPER identified by restriction digestion,the right vectors an lung adenocarcinoma cell line. The HER2 mRNA munofluorescence staining. FCM analysis and MTT method easure cell cycle and cell grober of cells in G0/G1 phase by %, and inhibit the tumor cell grober of cells in G0/G1 phase, and then sloor may be a useful therapeutic strategy for NSCLC overexpressing HER2/neu.
【Keya, nonsmallcell lung; HER2/neu
【摘要】目的:构建针对HER2/neu的RNA干涉载体,观察对HER2/neu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pSUPER质粒载体,转化DH5α菌株,提取质粒,转染过表达HER2/neu的肺腺癌细胞系SPCA1,RTPCR检测HER2/neu的mRNA表达, 分析细胞周期变化,MTT法观察细胞生长. 结果:肺腺癌细胞系SPCA1存在HER2/neu过表达;成功构建了HER2/neu特异性RNA干涉载体pSUPERsiHER2;瞬时转染SPCA1细胞,HER2/neu的mRNA及蛋白表达均降低; %;肿瘤细胞增殖速度减慢(P<). 结论:成功构建了HER2/neu RNA干涉载体,转染后可有效降低肺腺癌细胞系SPCA1 HER2/neu的mRNA转录及蛋白水平表达,阻滞转染细胞于G1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.
【关键词】 siRNA;RNA干涉;癌,非小细胞肺;HER2/neu
0引言

HER2/neu是由癌基因ErbB2/neu编码的具有酪氨酸蛋白激酶活性的跨膜糖蛋白,分子量为185 ku, 属表皮生长因子受体(epider