包载TIMP-1重组腺病毒微球的制备及其对肝癌细胞增殖的抑制  

Preparation of microsphere encapsulating recombinant TIMP-1 adenovirus and its inhibitory effects against hepatocellular carcinoma cells

作　　者：夏冬[1] 吴斌[1] 梁建群[1] 余少鸿[2] 徐亮[1] 

机构地区：[1]泸州医学院附属医院普外科,四川泸州646000 [2]重庆市涪陵中心医院肝胆外科,重庆400800

出　　处：《中国肿瘤生物治疗杂志》2010年第1期57-61,共5页Chinese Journal of Cancer Biotherapy

基　　金：四川省教育厅课题资助(No.2006B108);四川省卫生厅课题资助(No.090210)~~

摘　　要：目的:制备携带人基质金属蛋白酶组织抑制因子-1(tissue inhibitors of metalloproteinase-1,TIMP-1)的重组腺病毒乳酸聚乙烯醇(poly-DL-lactide-poly,PELA)微球,探讨其对HepG2肝癌细胞增殖的影响。方法:采用溶剂挥发法双乳液体系,以可降解的生物材料PELA包被携带TIMP-1基因的重组腺病毒制成微球,测定其粒径、载病毒量、包封率及释放规律。重组腺病毒微球感染HepG2细胞,荧光显微镜观测感染效率,透射电镜观测超微结构,半定量RT-PCR检测TIMP-1mRNA表达;MTT法检测HepG2细胞增殖。结果:成功构建包载TIMP-1重组腺病毒的PELA微球,直径约1.965μm,包封率为60%,载病毒率为10.5×108efu/mg,在120h内释放病毒量接近60%,总的释放时间长于240h。空白微球无毒性PELA病毒微球感染HepG2细胞后,细胞稳定表达TIMP-1mRNA;对HepG2细胞的增殖有明显抑制作用,抑制率表达47%。结论:包载TIMP-1重组腺病毒的PELA微球可抑制肝癌HepG2细胞的增殖,为化学高分子载体运载基因治疗肝癌提供了实验依据。Objective：To prepare poly-DL-lactide-poly （PELA） microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 （TIMP-1） adenovirus,and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells.Methods：The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA.The diameter of the microsphere,quantity of virus encapsulated,loading rate,and releasing kinetics were measured.HepG2 cells were infected with the microspheres;the infection efficiency was examined by fluorescent microscope;and the ultrastructure was observed by TEM.The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR,and the proliferation of HepG2 cells was detected by MTT assay.Results：The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed,with its diameter,entrapment efficiency,and virus loading rate being 1.965,60.0%,and 10.5×108/mg,respectively.About 60% of the viruses were released within 120 h,and the total releasing time was longer than 240 h.Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells,and significantly inhibited the proliferation of HepG2 cells,with the inhibitory rate being 47%.Conclusion：PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells,which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

关 键 词：肝肿瘤 基质金属蛋白酶组织抑制因子 腺病毒 微球 基因治疗 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]