Amifostine has antiangiogenic properties in vitro by changing the redox status of human endothelial cells.
Amifostine is a broad-spectrum cytoprotective agent, selective for normal tissues. It is a pro-drug metabolised to the free thiol WR-1065 that may act as a scavenger of free radicals, generated in tissues exposed to chemotherapeutic agents or radiation. WR-1065 can be further oxidized to its symmetric disulfide WR-33278 or degraded to hydrogen peroxide (H2O2). Both WR-1065 and WR-33278 resemble endogenous polyamines. Although amifostine is used in some cases in the clinic, there are only few studies concerning its actions at the cellular level. We have previously shown that amifostine inhibits angiogenesis in vivo, affecting the expression of several angiogenic genes. In the present work, we studied the effect of amifostine on human umbilical vein endothelial cell (HUVEC) functions in vitro, in order to further clarify its mechanism(s) of action. Amifostine increased HUVEC proliferation, an effect that was reversed by the intracellular H2O2 scavenger sodium pyruvate, agents that increase intracellular cAMP levels and L-valine. On the other hand, amifostine decreased HUVEC migration, an effect that was reversed by L-valine or L-arginine but not sodium pyrouvate. The decrease in migration was in line with decreased tube formation on matrigel and decreased amounts of metalloproteinase-2 released into the culture medium of HUVEC. Finally, amifostine reduced tyrosine nitration of the cytoskeletal proteins actin and alpha-tubulin in a time dependent manner. This last action could be due to the reduced production of nitric oxide (NO) or to other not yet identified mechanisms. Collectively, our results suggest that amifostine acts on endothelial cells through pathways that affect the redox status of the cells, either by producing H2O2 or by modulating NO production.
Giannopoulou
Efstathia
E
Department of Pharmacy, Laboratory of Molecular Pharmacology, University of Patras, GR 26504 Patras, Greece.
Papadimitriou
Evangelia
E
D000199:Actins / Q000378:metabolism; D004999:Amifostine / Q000378:metabolism / Q000494:pharmacology*; D020533:Angiogenesis Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen; D000242:Cyclic AMP / Q000378:metabolism; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007797:Laminin; D045726:Metalloproteases / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D010084:Oxidation-Reduction; D011509:Proteoglycans; D019289:Pyruvic Acid / Q000494:pharmacology; D014404:Tubulin / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
D000199:Actins; D020533:Angiogenesis Inhibitors; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D014404:Tubulin; C059485:matrigel; D009569:Nitric Oxide; D014443:Tyrosine; D019289:Pyruvic Acid; D003094:Collagen; D000242:Cyclic AMP; D045726:Metalloproteases; D004999:Amifostine