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Vanadate protects human neuroblastoma SH-SY5Y cells against peroxynitrite-induced cell death
Makio Saeki1, Sadaaki Maeda, Yoshinori Kamisaki
1Department of Pharmacology, Graduate School of Dentistry, Osaka University, Osaka, Japan. msaeki@dent.osaka-u.ac.jp
Abstract:
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).
Insights
Vanadate, a tyrosine phosphatase inhibitor, protects human neuroblastoma cells from peroxynitrite-induced death. This protection involves enhancing tyrosine phosphorylation of proteins like p130(cas), suggesting disrupted signaling is key to cell death.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Peroxynitrite is implicated in neuronal cell death.
- Tyrosine phosphorylation plays a role in cellular signaling pathways.
- Understanding cell death mechanisms is crucial for neurodegenerative disease research.
Purpose of the Study:
- To investigate the protective effect of vanadate against peroxynitrite-induced cell death in human neuroblastoma SH-SY5Y cells.
- To elucidate the role of tyrosine phosphorylation in peroxynitrite-induced cell death.
- To determine the signaling pathways involved in vanadate's protective mechanism.
Main Methods:
- Human neuroblastoma SH-SY5Y cells were treated with peroxynitrite donors (SIN-1) and inhibitors.
- Vanadate, okadaic acid, cyclosporin A, nitric oxide donors, and wortmannin were used.
- Protein tyrosine phosphorylation levels and protein-protein interactions (p130(cas)-Crk) were analyzed.
Main Results:
- Vanadate protected cells from SIN-1-induced death, unlike other phosphatase inhibitors.
- Vanadate's protective effect was independent of PI3-kinase.
- Vanadate increased tyrosine phosphorylation of p130(cas) and restored its association with Crk disrupted by SIN-1.
Conclusions:
- Disruption of tyrosine phosphorylation signaling is critical for peroxynitrite-induced cell death.
- Vanadate confers protection by enhancing tyrosine phosphorylation of proteins, including p130(cas).
- Vanadate represents a potential therapeutic strategy for conditions involving peroxynitrite-mediated neurotoxicity.