Vanadate protects human neuroblastoma SH-SY5Y cells against peroxynitrite-induced cell death.
We investigated the effect of vanadate, a tyrosine phosphatase inhibitor, on cell death induced by peroxynitrite in human neuroblastoma SH-SY5Y cells. Vanadate prevented cell death induced by 3-morpholinosydnonimine (SIN-1), a peroxynitrite donor; whereas SIN-1-induced cell death was not prevented by neither okadaic acid, an inhibitor of serine/threonine phosphatases 1 and 2A, nor cyclosporin A, an inhibitor of serine/threonine phosphatase 2B. Vanadate did not prevent cell death induced by N-ethyl-2-(1-ethyl-hydroxy-2-nitrosohydrazino)-ethanamine, a nitric oxide donor. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3-kinase), did not block the protective effect of vanadate, suggesting that the protective effect of vanadate is independent on PI3-kinase. Vanadate increased tyrosine phosphorylation of several proteins including the focal adhesion protein p130 Crk-associated substrate (p130(cas)). By the treatment with SIN-1, the endogenous association of p130(cas) and Crk was disrupted, and the association was restored by vanadate treatment. These results suggest that disruption of tyrosine phosphorylation signaling may be critical for peroxynitrite-induced cell death, and that vanadate prevents cell death at least in part through the enhancement in tyrosine phosphorylation of the proteins including p130(cas).
Saeki
Makio
M
Department of Pharmacology, Graduate School of Dentistry, Osaka University, Osaka, Japan. msaeki@dent.osaka-u.ac.jp
Maeda
Sadaaki
S
Kamisaki
Yoshinori
Y
D016923:Cell Death / Q000187:drug effects* / Q000502:physiology; D050719:Crk-Associated Substrate Protein; D016572:Cyclosporine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D008981:Molsidomine / Q000031:analogs & derivatives / Q000494:pharmacology; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D019319:Okadaic Acid / Q000494:pharmacology; D030421:Peroxynitrous Acid / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010750:Phosphoproteins / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors; D011506:Proteins; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D050737:Proto-Oncogene Proteins c-crk; D050717:Retinoblastoma-Like Protein p130; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology*
D050719:Crk-Associated Substrate Protein; D016572:Cyclosporine; D004791:Enzyme Inhibitors; D008981:Molsidomine; D020030:Nitric Oxide Donors; D019319:Okadaic Acid; D030421:Peroxynitrous Acid; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010750:Phosphoproteins; D017027:Protein Tyrosine Phosphatases; D011506:Proteins; D011518:Proto-Oncogene Proteins; D050737:Proto-Oncogene Proteins c-crk; D050717:Retinoblastoma-Like Protein p130; D014638:Vanadates; C493841:BCAR1 protein, human; C002385:linsidomine