靶向表皮生长因子受体的shRNA对实验性肝癌的治疗作用  

EGFR-targeted shRNA gene therapy for experimental human liver cancer mediated by PILP

作　　者：胡玉荣[1] 赵宏巧[1] 耿鑫[1] 季宝芳[1] 李春明[1] 张云[1] 张振中[1] 

机构地区：[1]郑州大学药学院,郑州450001

出　　处：《中国新药杂志》2013年第13期1513-1518,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金(81272563);河南省医学科技攻关项目(200803015)

摘　　要：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 effect of EGFR-targeted RNA interference treatment for liver canc- er. nethods：TR-PCR was used to detect the hEGFR genetic level in SMMC-7721 cells transfected in vitro. Cell cycle distribution and apoptotic rate were evaluated by flow cytometry （ FCM）. Nude mouse model was established. Each nude mouse was intravenously given 40 Ixg PILP-mediated anti-hEGFR pDNA. The mice were sacrificed after continuous administration for 5 times. The inhibition rate of tumor growth was calculated and the hEGFR protein ex- pression in tumor tissue was detected with Western Blot, and the downregulation of the hEGFR mRNA was detected by RT-PCR. Results： The PILP-mediated anti-hEGFR pDNA triggered hEGFR sequence-specific gene silencing effect with time-effect relationship. The results of flow cytometry cell cycle test and apoptosis test were consistent. In vitro, the anti-hEGFR pDNA targeted gene induced the apoptosis of hepatoma SMMC-7721 cells and inhibited the expression of EGFR significantly. In vivo, the EGFR expression of anti-hEGFR pDNA group was reduced by 56.72% , and the growth of subcutaneous tumor of hepatoma SMMC-7721 was inhibited significantly with inhibition rate of 46. 13% （P 〈 0.05）. Conclusion： PILP-mediated hEGFR mRNA induced cell apoptosis in SMMC-7721 cells, and inhibited tumor growth and down-regulated hEGFR protein production in tumor tissue in mice.

关 键 词：基因治疗 人表皮生长因子受体 RNA干扰 肝癌 

分 类 号：R966[医药卫生—药理学]