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目的:考察靶向人表皮生长因子受体(hEGFR)mRNA的RNA干扰(RNAi)对实验性肝癌的治疗作用。方法:制备聚乙二醇(PEG)修饰的靶向hEGFR mRNA的可表达短发夹RNA(shRNA)质粒的免疫脂质复合物(PILP);RT-PCR法检测体外转染的SMMC-7721细胞中hEGFR基因表达水平;流式细胞术检测细胞周期分布和凋亡。建立裸鼠移植瘤模型,计算肿瘤生长抑制率,检测肿瘤组织的EGFR基因和蛋白表达水平的变化。结果:PILP介导的anti-hEGFR pDNA具有引发hEGFR序列特异性基因沉默效应,且呈时间-效应关系。anti-hEGFR pDNA在体外诱导肝癌SMMC-7721细胞凋亡作用显著,并显著抑制该细胞中EGFR基因的表达。anti-hEGFR pDNA在体内可将肿瘤组织中EGFR表达降低56.72%,并显著抑制裸鼠皮下移植瘤生长,肿瘤生长抑制率达46.13%(P<0.05)。结论:PILP介导的靶向hEGFR mRNA的治疗质粒对实验性肝癌具有治疗作用。
Objective: To investigate the therapeutic effect of RNA interference targeting human epidermal growth factor receptor (hEGFR) mRNA on experimental liver cancer. METHODS: Immunoliposomes (PILP) expressing polyethylene glycol (PEG) -mediated shRNA targeting hEGFR mRNA were prepared and transfected into SMMC-7721 cells by RT-PCR In hEGFR gene expression levels; flow cytometry to detect cell cycle distribution and apoptosis. The model of xenografted nude mice was established, the growth inhibition rate was calculated, and the changes of EGFR gene and protein expression in tumor tissues were detected. Results: PILP-mediated anti-hEGFR pDNA had a hEGFR sequence-specific gene silencing effect and showed a time-effect relationship. Anti-hEGFR pDNA significantly induced apoptosis of SMMC-7721 cells in vitro and significantly inhibited the expression of EGFR gene in this cell line. Anti-hEGFR pDNA can reduce the EGFR expression in tumor tissue by 56.72% in vivo and significantly inhibit the growth of subcutaneous xenograft in nude mice. The inhibition rate of tumor growth was 46.13% (P <0.05). CONCLUSIONS: PILP -mediated therapeutic plasmid targeting hEGFR mRNA has a therapeutic effect on experimental liver cancer.