The Experimental Study On Oncolytic Recombinant Adenovirus Ad-E1A Inhibit A375 In Vitro And H22 Tumor Growth In Vivo

Objective:To construct The oncolytic adenovirus Ad-E1A and explore the effect of E1A on A375 cells.It inhibit H22 tumor growth in vivo and enhance the immune function of BALB/c mice.Methods:The pAdTrack-CMV-E1A was constructed with pUCm-T-E1A recombined plasmid by PCR as template,enzyme digestion and ligation.The pAdTrack-CMV-E1A lineared by PmeI was co-transducted into BJ5183 with pAdeasy-I.Analysis of PCR and DNA sequencing was carried out to demonstrate the sequence of the piasmid.The pAdeasy-1-pAdTrack-CMV-E1A recombined adenovirus vector was leared with PacI and then transfected into QBI-293A cells.The Ad-E1A recombined adenovirus was obtained and was used to infect A375 cells.Identification of Ad-E1A mRNA by RT-PCR was carried out to demonstrate the expression of Ad-E1A in A375 cells.The effect of Ad-E1A on A375 was studied by MTT,flow cytometry.And study the effect of Ad-E1A on H22 tumor growth in vivo and the immune function of BALB/c mice.Results:The recombined oncolytic adenovirus Ad-E1A was obtained. The oncolytic adenovirus inhibit H22 ascitic hepatoma obviously,have no side effect on immune organ and enhance the numbers of monocyte and neutrophil. Conclusion:The oncolytic adenovirus Ad-E1A is constructed succefully.The oncolytic adenovirus Ad-E1A inhibites A375 obviously.It inhibit H22 tumor growth in vivo and enhance the immune function of BALB/c mice.