肺腺癌SPC-A-1细胞表皮生长因子受体表达水平对其增殖的影响  

Antitumor effect of epidermal growth factor receptor gene silencing

作　　者：张敏[1] 白春学[1] 张新[1] 高磊[1] 毛翎[1] 陈杰[1] 

机构地区：[1]复旦大学附属中山医院呼吸病研究所呼吸内科,上海200032

出　　处：《中华结核和呼吸杂志》2005年第5期337-341,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：上海市科委重大科技攻关项目(04DZ19208);上海市科技发展基金资助项目(03ZR14004)

摘　　要：目的探讨采用体外化学合成的针对表皮生长因子受体(EGFR)设计的双链RNA(doublestrandedRNA,dsRNA),是否能诱导肺非小细胞肺癌(NSCLC)细胞出现序列特异性基因沉默,及抑制EGFR基因表达后NSCLC细胞的生物学特征。方法体外合成EGFR序列特异性dsRNA(dsRNAEGFR),结合脂质体2000转染肺腺癌SPC-A-1细胞,用荧光显微镜及流式细胞仪测定转染细胞的EGFR受体数量;适时定量聚合酶链反应检测其EGFR基因表达水平。采用集落形成试验检测SPC-A-1细胞的增殖和集落形成能力。建立裸鼠肿瘤模型,计算肿瘤抑制率。结果dsRNAEGFR可使EGFR在蛋白水平表达下降71.3%、基因水平表达下降50.0%,SPC-A-1细胞集落形成下降66.8%,并显著抑制在体肿瘤生长,肿瘤抑制率为75.0%。结论dsRNAEGFR可序列特异性下调NSCLC细胞EGFR在基因及蛋白水平的表达,有效抑制肿瘤生长。Objective To investigate whether chemically synthesized double-stranded RNA(dsRNA) targeting epidermal growth factor receptor(EGFR) could induce gene silencing in non-small-cell lung cancer (NSCLC) cells, and to assess the degree of EGFR gene silencing and its effect on functional outcome. Methods NSCLC cell line SPC-A-1 was transfected with target sequence-specific dsRNA formulated with Lipofectamine 2000. Fluorescent microscopy and flow cytometry were used to measure the reduction in the production of the EGFR protein. Real-time PCR was used to detect the silencing of the EGFR gene level. Colony assay was adopted to measure the cellular proliferation and colony formation. A tumor burdened athymic nude mouse model was established to calculate the tumor growth inhibition rate. Results The dsRNA-EGFR was shown to be effective with a 71.3% down-regulation of EGFR protein production and 50.0% of silencing of EGFR gene. The dsRNA-EGFR significantly reduced colony numbers by 66.8% in vitro and inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.0%. Conclusion The sequence specific dsRNA showed a blockbuster effect in downregulation of EGFR gene level and protein production, inhibition of the cellular proliferation and tumor growth.

关 键 词：SPC-A-1细胞 表皮生长因子受体表达 肺腺癌 表皮生长因子受体(EGFR) 非小细胞肺癌(NSCLC) 增殖 聚合酶链反应检测 序列特异性 肿瘤抑制率 EGFR受体 基因表达水平 集落形成能力 集落形成试验 细胞集落形成 抑制肿瘤生长 

分 类 号：R734.2[医药卫生—肿瘤] R739.41[医药卫生—临床医学]