Preliminary effect of VEGF promoter-driven recombinant adenovirus containing double suicide genes on apoptosis of human gastric carcinoma cells.
- Author:
Heng KONG
1
;
Zong-Hai HUANG
;
Zhou LI
;
Jing-Long YU
;
Hai-Jin CHEN
;
Xin-Jun HAN
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; physiology; Animals; Apoptosis; drug effects; genetics; Calcium; metabolism; Cell Line, Tumor; DNA; metabolism; DNA, Recombinant; genetics; Dose-Response Relationship, Drug; Flucytosine; pharmacology; Ganciclovir; pharmacology; Genes, Transgenic, Suicide; genetics; Humans; Microscopy, Electron; Promoter Regions, Genetic; genetics; Stomach Neoplasms; genetics; metabolism; pathology; virology; Vascular Endothelial Growth Factor A; genetics
- From: Journal of Southern Medical University 2007;27(8):1152-1160
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSVEGF-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.
RESULTSThe 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.
CONCLUSIONSCD/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.