Selective killing of vascular endothelial cells based on adenovirus-mediated herpes simplex virus thymidine kinase gene transfection under the driving of KDR promoter

AIM: To investigate the target killing effect of adenovirus-mediated herpes simplex virus thymidine kinase gene(HSV-tk) transfection under the driving of KDR promoter on the vascular endothelial cells in vitro.METHODS:By using AdEasy system,recombinant adenovirus plasmid containing KDR or cytomegalovirus(CMV) promoter-controlled HSV-tk gene(AdKDR-tk and AdCMV-tk) was constructed.After packaging and amplification in 293 cells,the virus was used to infect KDR-expressed human umbilical venous endothelial cells(HUVEC) and KDR-unexpressed CNE-2.Following administration of ganciclovir(GCV),the survival rate of gene-transfected HUVEC and CNE-2 was evaluated by using MTT method.RESULTS:The AdEasy System produced a high titer of the recombinant adenovirus(1×10~(13) pfu/L).Under infection index of 100,with increasing GCV concentration from 0 up to 50 mg/L, the survival rates of AdKDR-tk-transfected HUVEC and CNE-2 decreased from(89.4±(4.6)%) and(91.5±4.4)% to(22.9±4.7)% and(71.4±(2.9)%) at proper order respectively(P0.01),while the(survival) rates of AdCMV-tk-transfected HUVEC and CNE-2 declined from(89.9±6.2)% and(90.8±5.7)% to(12.8±(2.6)%) and(18.8±6.1)%,respectively(P0.05).(CONCLUSION:)Adenovirus-mediated HSV-tk transfection under the driving of KDR promoter could yield the specific killing effect on vascular endothelial cells with treatment of GCV.