Mechanism of recombinant adenovirus-mediated mutations of hypoxia inducible factor 1alpha in modulation of cell apoptosis.
- Author:
Li-li WEI
1
;
Ping-sheng WU
;
Yue-gang WANG
;
Ying-fang HU
;
Yi-jun XIE
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; genetics; Apoptosis; genetics; physiology; Blotting, Western; Cell Line, Tumor; Colonic Neoplasms; genetics; metabolism; pathology; Cyclin-Dependent Kinase Inhibitor p21; biosynthesis; genetics; Genetic Vectors; genetics; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; biosynthesis; genetics; Mutagenesis, Site-Directed; Mutation; RNA, Messenger; biosynthesis; genetics; Recombinant Proteins; biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; Transfection
- From: Journal of Southern Medical University 2008;28(3):309-312
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the mechanism by which recombinant adenovirus (Ad)-mediated mutations of hypoxia inducible factor 1alpha (Ad-HIF-1alpha-Ala564-Ala803) regulates cell apoptosis.
METHODSLoVo cells were infected with recombinant Ad-HIF-1alpha-Ala564-Ala803 and control virus Ad-lacZ under normoxia condition. Real-time PCR was used to detect HIF-1alpha and p21WAF1/CIP1 mRNA expressions at different time points. Western blotting was employed to verify HIF-1alpha and p21WAF1/CIP1 protein expression. Hoechst 33342 flourescein staining was performed to observe the ratio of apoptotic LoVo cells.
RESULTSThe expression levels of HIF-1alpha mRNA and protein increased after infection with Ad-HIF-1alpha- Ala564-Ala803, accompanied by an increase in p21WAF1/CIP1 mRNA and protein expressions. The apoptotic ratio was significantly higher in LoVo cells infected with recombinant Ad-HIF-1alpha-Ala564-Ala803 (16.2%) than in the control cells (5.5%, P=0.00).
CONCLUSIONHIF-1alpha may induce cell cycle arrest by up-regulating p21WAF1/CIP1 expressions to promote LoVo cell apoptosis.
