Gene transfer of CuZn superoxide dismutase enhances the synthesis of vascular endothelial growth factor.
Nitric oxide (NO) and reactive oxygen species (ROS) are emerging as important regulators of angiogenesis. NO enhances VEGF synthesis in several cell types and is required for execution of VEGF angiogenic effect in endothelial cells. Similarly, hydrogen peroxide induces VEGF synthesis and recent studies indicate the involvement of ROS in signaling downstream of VEGF stimulation. VEGF synthesis can not only be enhanced by gene transfer of VEGF but also by overexpression of NO synthase genes. Here, we examined the possibility of augmentation of VEGF production by gene transfer of copper/zinc superoxide dismutase (CuZnSOD, SOD1). Overexpression of human SOD1 in mouse NIH 3T3 fibroblasts increased SOD activity, enhanced intracellular generation of H2O2 and significantly stimulated VEGF production as determined by increase in VEGF promoter activity, VEGF mRNA expression and VEGF protein synthesis. The stimulatory effect on VEGF synthesis induced by SOD1 gene transfer was reverted by overexpression of human catalase. The effect of H2O2 produced by engineered cells is mediated by activation of hypoxia-inducible factor response element (HRE) as well as Sp1 recognition site of VEGF promoter. This data suggest the feasibility of stimulation of angiogenesis by overexpression of SOD1.
Grzenkowicz-Wydra
Jolanta
J
Department of Cell Biochemistry, Faculty of Biotechnology, Jagiellonian University, Kraków, Poland.
Cisowski
Jarosław
J
Nakonieczna
Joanna
J
Zarebski
Adrian
A
Udilova
Natalia
N
Nohl
Hans
H
Józkowicz
Alicja
A
Podhajska
Anna
A
Dulak
Józef
J
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002374:Catalase / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D041681:NIH 3T3 Cells; D018919:Neovascularization, Physiologic; D009569:Nitric Oxide / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017382:Reactive Oxygen Species / Q000378:metabolism; D020218:Response Elements; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D016329:Sp1 Transcription Factor / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics*; D013481:Superoxides / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D017382:Reactive Oxygen Species; D016329:Sp1 Transcription Factor; D042461:Vascular Endothelial Growth Factor A; D013481:Superoxides; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D001616:beta-Galactosidase