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Inhibitory effect of 5/35 chimeric oncolytic adenovirus SG635 on hepatocarcinoma cells
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Abstract:
Objective : To investigate the specific cytotoxicity effect of 5/35 chimeric oncolytic adenovirus SG635 on hepatocellular carcinoma HepG2 and SMMC-7721 cells. Methods: The knob and shaft domains of type 5 adenovirus (Ad5) in SG600 plasmid were replaced by the domains of type 35 adenovirus (Ad35), and chimeric oncolytic adenovirus Ad5/35 was established. Flow cytometry was used to examine the infection efficiency of chimeric adenovirus Ad5/35 (Ad5/35-EGFP) in HepG2 and SMMC-7721 cells; replication assay was used to evaluate the replication of oncolytic adenovirus SG635; Western blotting analysis was used to examine the expression of E1A in cells after SG635 infection; and Kit-8 assay was used to assess the cytotoxicity of SG635 and SG600 on HepG2 and SMMC-7721 cells. Results: The infection efficiency of Ad5/35-EGFP in HepG2 and SMMC-7721 cells was obviously enhanced compared with Ad5-EGFP. The replication activity of SG635 in HepG2 and SMMC-7721 cells was higher than that of SG600 72 h after infection (15 848.93, 6 309.57 vs 6 309.57, 5 011.87, P<0.01), but SG635 did not replicate in normal BJ cells. Moreover, SG635 induced a higher expression of E1A protein in HepG2 and SMMC-7721 cells than SG600, but did not induce E1A expression in normal BJ cells. At a certain MOI, SG635 showed increasing cytotoxicity on HepG2 (MOI=1, 90% vs 60%) and SMMC-7721 (MOI=10, 90% vs 50%) cells, and the cytotoxicity was stronger than SG600, without causing significant cytotoxicity on normal BJ cells. Conclusion: The 5/35 chimeric oncolytic adenovirus SG635 can effectively infect and specifically kill hepatocarcinoma cells with satisfactory safety and specificity.
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Project Supported:
Project supported by the State Key Foundation of Science and Technology Ministry during the"11th Five-Year Plan" of China (No. 2008ZX10002-025, No. 2008ZX10002-026), the Special Foundation for State Major Basic Research Program of China (973 program) (No. 2009CB522404), the Foundation for New Century Excellent Talents of Ministry of Education of China (No. NCET-08-0583), and the Foundation for the Author of National Excellent Doctoral Dissertation of China (No. FANEDD 200774)
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