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1 mRNA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1.965 μm，包封率为60%，载病毒率为10.5×108efu/mg，在120 h内释放病毒量接近60%，总的释放时间长于240 h。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1 mRNA；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 10.5×108/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.

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