[A double suicide gene system driven by KDR promoter selectively kills human hepatic carcinoma cells and human umbilical vein endothelial cells].
OBJECTIVE
To study the selective killing effects of adenovirus (Ad)-mediated double suicide gene system driven by KDR promoter (KDR-CdglyTK) on the human hepatic carcinoma cells and human umbilical vein endothelial cells (HUVECs).


METHODS
KDR-expressing BEL-7402 and HUVECs and HepG2 cells that did not express KDR were infected by KDR-CdglyTK, and the infection efficiency and the expression of CdglyTK in the cells was detected by RT-PCR. The infected cells were treated with the the prodrugs 5-FC and GCV at different concentrations, and the cell-killing effects and bystander effects were evaluated by MTT method.


RESULTS
At the multiplicity of infection (MOI) of 100, the recombinant AdKDR-CDglyTK showed similar infection efficiency in the 3 cell lines. RT-PCR demonstrated CDglyTK expression in the recombinant adenovirus and the 3 infected cell lines. BEL-7402 and HUVECs infected by the KDR-CdglyTK, but not the HepG2 cells, were highly sensitive to the prodrugs (P<0.001). Bystander effects of the double suicide gene system were observed in the coculture of the infected and non-infected BEL-7402 and HUVECs.


CONCLUSIONS
The double suicide gene system driven by KDR promoter has specific killing effect on KDR-expressing hepatocellular carcinoma cells and HUVECs.
Yang
Liu-cheng
LC
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. sdylc@yahoo.com.cn
Huang
Zong-hai
ZH
Kong
Heng
H
Li
Qiang
Q
Chen
Fei
F
Yu
Jin-long
JL
Li
Zhou
Z
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis / Q000235:genetics; D002478:Cells, Cultured; D043525:Cytosine Deaminase / Q000235:genetics* / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014471:Umbilical Veins / Q000166:cytology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D013937:Thymidine Kinase; D040301:Vascular Endothelial Growth Factor Receptor-2; D043525:Cytosine Deaminase