检索历史 清除

目的 探讨体外化学合成表皮牛长因子受体(EGFR)基因序列特异性双链RNA(dsRNA)在体内诱导非小细胞肺癌(NSCLC)细胞出现序列特异性基因沉默的町行性.方法 体外化学合成EGFR序列特异性dsRNA(dsRNA-EGFR),结合脂质体Lipofectamine 2000转染肺腺癌细胞株SPC-A1后,将200 μl细胞悬液接种于裸鼠,建立荷瘤鼠模型,计箅肿瘤抑制率.采用免疫组织化学技术、Western blot技术和实时逆转录聚合酶链反应(real-time RT-PCR),检测肿瘤组织中EGFR蛋白和mRNA的表达水平.结果 dsRNA-EGFR可显著抑制体内肿瘤生长,肿瘤抑制率为75.0%,并可将EGFR蛋白表达水平降低53.6%、mRNA表达水平降低32.3%.结论 dsRNA-EGFR在体内可有效抑制NSCLC细胞中EGFR蛋白和mRNA的表达水平,抑制肿瘤生长.

Objective To investigate whether chemically synthesized double-stranded RNA (dsRNA)targeting epidermal growth factor receptor(EGFR)can induce gene silencing in non-small cell lung cancer(NSCIJC)cells in vivo.Methods The NSCLC cellJine SPC.Al was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000.SPC-Al cells(1×107/ml)in 200 μl were injected s.c.into the left flank area of the athymic nude mice to establish a tumor-bearing nude mouse model.To calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor.Immunohistochemistry and Western blot were used to monitor the reduction of EGFR protein production.Real-time RT-PCR Was used to detect the silencing of the EGFR mRNA level.Results The EGFR sequence specific dsRNA(dsRNA-EGFR)significantly inhibited the tumor growth in vivo.The tumor growth inhibition rate was 75.0%.The dsRNA-FGFR sequence specifically silenced EGFR with 53.6%of downregulation of EGFR protein production and 32.3%of silencing of EGFR mRNA level.Conclusion dsRNAEGFR show a blockbuster effect in down-regulation of EGFR mRNA leveI and protein production.and inhibition of tumor growth in vivo.