Abstract
We have isolated a gene, the c subunit (ATP6L) of vacuolar H(+)-ATPase, involved in oxidative stress response. In this study, we examined the role of ATP6L and its molecular mechanisms in glial cell death induced by H(2)O(2). Expression of the ATP6L gene was increased by H(2)O(2) treatment in C6 glial cells. ATP6L siRNA-transfected C6 cells treated with H(2)O(2) showed a significant decrease in viability. ATP6L siRNA-transfected cells that were pretreated with MEK1/2 inhibitor completely recovered cell viability. Pretreatment of the transfected cells with zVAD-fmk, a pan-specific caspase inhibitor, did not result in the recovery of cell viability, as determined by a H(2)O(2)-induced cytotoxicity assay. The ultrastructural morphology of the transfected cells as seen by the use of transmission electron microscopy showed numerous cytoplasmic autophagic vacuoles with double membrane. These results suggest that ATP6L has a protective role against H(2)O(2)-induced cytotoxicity via an inhibition of the Erk1/2 signaling pathway, leading to inhibition of autophagic cell death.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Autophagy / drug effects
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Autophagy / physiology
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Brain / enzymology*
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Brain / physiopathology
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Cell Death / drug effects
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Cell Death / physiology
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Cell Line, Tumor
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Cell Survival / drug effects
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Cell Survival / physiology
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Cytoprotection / drug effects
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Cytoprotection / physiology
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Enzyme Inhibitors / pharmacology
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Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
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Extracellular Signal-Regulated MAP Kinases / metabolism
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Glioma
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Hydrogen Peroxide / toxicity*
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MAP Kinase Kinase 1 / antagonists & inhibitors
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MAP Kinase Kinase 1 / metabolism
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Microscopy, Electron, Transmission
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Neuroglia / drug effects
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Neuroglia / enzymology*
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Oxidants / toxicity
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Oxidative Stress / drug effects
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Oxidative Stress / physiology*
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Protein Subunits / genetics
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Protein Subunits / metabolism
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RNA, Small Interfering
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Rats
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Transfection
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Vacuolar Proton-Translocating ATPases / genetics
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Vacuolar Proton-Translocating ATPases / metabolism*
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Vacuoles / enzymology
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Vacuoles / ultrastructure
Substances
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Enzyme Inhibitors
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Oxidants
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Protein Subunits
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RNA, Small Interfering
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Hydrogen Peroxide
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Extracellular Signal-Regulated MAP Kinases
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MAP Kinase Kinase 1
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Vacuolar Proton-Translocating ATPases