目的：制备携带人基质金属蛋白酶组织抑制因子1（tissue inhibitors of metalloproteinase1，TIMP1）的重组腺病毒乳酸聚乙烯醇（polyDLlactidepoly，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TIMP1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RTPCR检测TIMP1 mRNA表达；MTT法检测HepG2细胞增殖。结果：成功构建包载TIMP1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10.5×108efu/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TIMP1 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TIMP1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 polyDLlactidepoly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase1 (TIMP1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods:The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP1 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 TIMP1 mRNA in HepG2 cells was examined by semiquantitative RTPCR, and the proliferation of HepG2 cells was detected by MTT assay. Results:The microsphere encapsulating recombinant TIMP1 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 rAdTIMP1 PELA microsphere efficiently induced TIMP1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion:PELA microsphere encapsulating recombinant TIMP1 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.