Recombinant adenovirus-mediated vascular endothelial growth factor gene transfer attenuates hypoxia-induced apoptosis of neural stem cells in vitro.
OBJECTIVE
To study the effects of vascular endothelial growth factor (VEGF) gene transfer on hypoxia-induced apoptosis of neural stem cells in vitro.


METHODS
C17.2 neural stem cells cultured in vitro were infected by recombinant adenovirus containing VEGF gene and cultured under hypoxic condition. VEGF expression in these cells was detected by Western blotting, and the apoptotic index was calculated from results of triphosphate-biotin nick end-labeling (TUNEL) assay. Flow cytometry was employed to examine the changes in the cell apoptotic rate after VEGF gene transfer, and the apoptotic bodies were observed under fluorescence microscope with Hoechst33342 staining.


RESULTS
The expression of VEGF was significantly increased in pAdCMV VEGF(165)-infected cells, resulting in inhibition of the apoptosis of C17.2 neural stem cells induced by hypoxia manifested by a significantly lower apoptotic rate of the stem cells transfected by pAdCMV VEGF(165) than that of the untransfected cells (10.38%;+/-0.48%; vs 19.98 %;+/-0.55%;, P<0.01) and of the cells transfected with pAdCMV VEGF(165) along with VEGF anti-sense oligodeoxynucleotide (19.07%;+/-0.64%;, <0.01) after hypoxia.


CONCLUSIONS
Recombinant adenovirus can efficiently mediate VEGF gene transfer into C17.2 neural stem cells, resulting in high expression of the exogenous VEGF in vitro, which effectively reduces C17.2 neural stem cell apoptosis induced by hypoxia.
Shen
Qing-Yu
QY
Department of Neurology, Second Affiliated Hospital, Sun Yat-sen University, Guangzhou 510120, China. shenqy@hotmail.com
Lü
Rui-Yan
RY
Li
Mei
M
Wang
Li-Mei
LM
Xiao
Song-Hua
SH
Xing
Yi-Gang
YG
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism*; D017209:Apoptosis / Q000502:physiology*; D002454:Cell Differentiation; D015687:Cell Hypoxia; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D009474:Neurons / Q000166:cytology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013234:Stem Cells / Q000166:cytology*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A