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Preparation of microsphere encapsulating recombinant TIMP-1 adenovirus and its inhibitory effects against hepatocellular carcinoma cells
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Abstract:
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.
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Project Supported:
Project supported by the Education Department Foundation of Sichuan Province (No.2006B108), and the Health Department Foundation of Sichuan Province (No.090210)
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