[Preliminary effect of VEGF promoter-driven recombinant adenovirus containing double suicide genes on apoptosis of human gastric carcinoma cells].
OBJECTIVE
To study the effect of the adenovirus containing CD/TK fusion gene controlled by the human vascular endothelial growth factor (VEGF) promoter on apoptosis of human gastric carcinoma cells SGC-7901.


METHODS
VEGF-expressing SGC-7901 cells were infected by the recombinant adenovirus Ad-VEGFP-CD/TK, and the infection efficiencies were observed with fluorescence microscopy. The toxic effect and intracellular calcium concentration induced by 5-fluorocytosine (5-FC) and ganciclovic (GCV) were determined by light microscopy, electron microscopy and flow cytometry.


RESULTS
The transfection efficiency of the recombinant adenovirus in SGC-7901 cells increased with the viral titer. At the multiplicity of infection (MOI) of 100, 5-FC and GCV could induce apoptosis of SGC-7901 cells within a given dose range in a dose- and time-dependent manner, and apoptotic changes of the cells were observed with electron microscopy. Apoptotic peak was also detected by flow cytometry. Cell cycle analysis revealed increased cell percentage in G(0)-G(1) phase and decreased percentage of cells in G(2)-M and S phases in response to treatment with the pro-drugs, which also induced marked elevation of intracellular calcium concentration in the infected cells.


CONCLUSIONS
CD/TK fusion gene system driven by VECF promoter selectively induces apoptosis of VEGF-expressing SGC-7901 cells, the action of which is probably mediated by intracellular calcium variation.
Kong
Heng
H
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China. generaldoc@ 126.com
Huang
Zong-Hai
ZH
Li
Zhou
Z
Yu
Jing-Long
JL
Chen
Hai-Jin
HJ
Han
Xin-Jun
XJ
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005437:Flucytosine / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D006801:Humans; D008854:Microscopy, Electron; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
D004274:DNA, Recombinant; D042461:Vascular Endothelial Growth Factor A; D004247:DNA; D005437:Flucytosine; D015774:Ganciclovir; D002118:Calcium