Construction of Adenovirus-mediated Ang1 and VEGF165 Co-expression Adenovirus Vector and Its Proangiogenesic Activity

Objective: To construct a recombinant adenoviral vector co-expressing angiogenin-1(Ang-1) and vascular endothelial growth factor(VEGF) and to provide experimental evidence for the effect of the transgenic fibroblasts on inducing angiogenesis.Methods: The co-expressing adenoviral vector Ad-Ang1-IRES-VEGF165 was constructed by cloned sequence and recombined technology.The recombinant adenoviruses were transfected into QBI-293A cells and tested it's titer.Then the WI-38 cells were transfected by the adenovirus Ad-AV and the expression of Ang1 and VEGF were identified by enzyme-labeled immunosorbent assay(ELISA).Meanwhile,the vessel forming test of chorioallantoic membrane(CAM) of enbryonated eggs was made to observe the growth of vessels.Results: The recombined adenovirus Ad-AV could be constructed and it was successfully packaged with the titer being as high as 4×1010 pfu/ml.The aim gene Ang-1 and VEGF165 could be effectively express in 293A cells and WI-38 cells.The CAM blood vessel of test group added with culture supernatantwere was more than that in control group(P0.01).Conclusion: The recombined adenovirus Ad-AV can be constructed successfully.Both Ang-1 and VEGF165 effectively express in target cells and the target gene had a marked proangiogenesic activity.