[Effect of KDR recombinant adenovirus containing double suicide gene on proliferation, apoptosis and cell cycle of human umbilical vein endothelial ECV304 cells].
OBJECTIVE
To study the effect of adenovirus (Ad)-mediated fusion gene systemdriven by KDR promoter on the proliferation, apoptosis and cell cycle of human umbilical vein endothelial ECV304 cells.


METHODS
The KDR-expressing ECV304 cells and LS174T cells not expressing KDR were both infected by the AdEasy-KDR-CDglyTK followed by treatment with the prodrugs 5-flurocytosine (5-FC) and/or ganciclovir (GCV) at different concentrations. The killing effects of the transfection on the cells were evaluated and bystander effects analyzed by coculturing the uninfected cells by AdKDR-CDglyTK with different ratios of infected cells. Flow cytometry was employed for determining the cell cycle distribution and electron microscopy performed to observe the pathological changes of cells.


RESULTS
The infection rates of the resultant recombinant Ad (rAd) were similar in the cells and gradually increased with the increment in the multiplicity of infection (MOI) of the Ads. The infected cells exhibited different sensitivities to the two prodrugs: ECV304 cells infected with rAd were highly sensitive to the prodrugs, but the infected LS174T cells were not (P<0.001). The killing effect of CD/TK fusion gene on the target cells was much stronger than that of either single suicide gene (P<0.001), showing also obvious bystander effect. In addition, the cell cycle of ECV304 cells was arrested at S phase with morphologic features of apoptosis and necrosis as displayed by electron microscopy.


CONCLUSIONS
CD/TK fusion gene system driven by KDR promoter selectively kills the KDR-CDglyTK-expressing endothelial cells, the mechanism of which may involve cell cycle arrest and necrosis and apoptosis of the cells.
Su
Guo-qiang
GQ
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. suguoqiang66@yahoo.com.cn
Huang
Zong-hai
ZH
Yu
Jin-long
JL
Li
Zhou
Z
Fan
Ying-fang
YF
Song
Hui-juan
HJ
Wang
Wei
W
Zhang
Jin-hua
JH
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis / Q000502:physiology*; D002453:Cell Cycle; D049109:Cell Proliferation; D002478:Cells, Cultured; D043525:Cytosine Deaminase / Q000235:genetics; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D020794:Receptor Protein-Tyrosine Kinases; D043525:Cytosine Deaminase