Construction of the oncolytic adenovirus Ad-E1A and its effect on hepatocellular carcinoma cell in vitro

Objective To construct the oncolytic adenovirus Ad-E1A and study its the cytolytic effect on human hepatocellular carcinoma(HCC) cell line.Methods The pAdTrack-CMV-E1A was constructed by PCR with QBI-293A genome DNA as template,enzyme digestion and ligation.The pAdTrack-CMV-E1A lineared by PmeI was co-transformed into BJ5183 with pAdEasy-1.The pAdEasy-1-pTrack-CMV-E1A recombined adenovirus vector was lineared with PacI and then transfected into QBI-293A cells.The E1A recombined adenovirus was obtained.The effect on cell growth and cell cycle of human HCC cell line(SMMC-7721) infected with Ad-E1A was determined based on cycotoxicity to SMMC-7721 by MTT assay and RT-PCR analysis.Results The pAdEasy-1-pTrack-CMV-E1A vector was constructed and the high titre E1A recombined adenovirus was obtained.Ad-E1A inhibited SMMC-7721 obviously,especially 24 hours after infection of tumor cells,and induced the SMMC-7721 apoptosis.Conclusion The oncolytic adenovirus Ad-E1A is constructed succefully,which inhibits SMMC-7721 obviously and induces the human hepatoma carcinoma cell apoptosis.