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Cytotoxicity of a novel replicative adenovirus CNHK500-hγ against hepatocellular carcinoma cells
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Abstract:
Objective : To investigate the cytotoxicity of the novel replicative adenovirus CNHK500-hγ, a recombinant adenovirus with the adenovirus E1A and E1B genes driven by human telomerase reverse transcriptase (hTERT) and hypoxia response element (HRE) promoters respectively and carrying the hIFN-γ, against hepatocellular carcinoma (HCC) cells in vitro. Methods: The amplification and cytotoxicity of replicating adenovirus CNHK500-hγ on two telomerase positive HCC cell lines (HepG2 and Hep3B) and one telomerase negative normal cell line (BJ) were analyzed by TCID50 and MTT assays. BJ, Hep3B and HepG2 cells were infected with CNHK500-GFP carrying green fluorescent protein and the amplification of CNHK500-GFP was observed. The expressions of hIFN-γ in cells and cell supernatants after CNHK500-hγ infection were detected by Western blotting and ELISA assays. Results: Forty-eight hours after infection, the amplification of CNHK500-GFP in HepG2 and Hep3B cells were 16 003 and 2 116 times of that in BJ cells, and the cytotoxicity ED50 of CNHK500-GFP against BJ cells was respectively 500 and 10 000 times of that against HepG2 and Hep3B cells, and superior to the positive control of replicative adenovirus ONYX-015. Furthermore, hIFN-γ expression in HepG2 and Hep3B cells after CNHK500-hγ infection was significantly higher than that after non-replicative adenovirus Ad-hγ infection (P<0.01). Conclusion: Replicative adenovirus CNHK500-hγ can specifically amplify in HCC cells and effectively express hIFN-γ gene, which holds potential for the treatment of HCC.
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Project Supported:
Project supported by the International Cooperation Important Project of National Natural Foundation of China (No. 30120160824), and the National High Science and Technology Research and Development Program of China (863 Program) (No.2001AA217031)
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