Construction of recombinant adenoviruse vector containing tissue-type plasminogen activator and its effect on the expression of VEGF in the transfected active ECV304 cells

Objective To investigate the effects of extrinsic tissue plasminogen activator(t-PA) on the expression of VEGF in ECV304 cells.Methods The adenovirus t-PA(Adt-PA) with the method of homologous recombination in bacterium was contrusted and transfected active ECV304 cells with different virus titers.The trasfection ratioes were 1∶10,1∶50 and 1∶100 and the fitting MOI value was selected.The expression of t-PA protein was detected with Western blot analysis after transfection.ECV304 cells were divided into three groups of A(Adt-PA cocultured for 24 hours),B(Adt-PA cocultured for 48 hours),and C(cultured with Ad for 48 hours as the control).The expressions of VEGF mRNA and VEGF protein were compared.Results Recombinant adenoviruse vector containing t-PA was contrasted successfully.Adt-PA had some promotion effects on proliferation of ECV304 at 1∶50,also the transfective efficiecy is(69.6±21.2)%.After transfection,the quantity of t-PA expression increased gradually within 72 hours(P0.01).The expressions of VEGF mRNA and protein were obviously higher in groups of A and B than those in group C(P0.01).Conclusion Recombinant adenoviruse vector containing t-PA could transfect ECV304 cells and promote its VEGF expression.