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PMID:12551919
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| Citation |
Pias, EK, Ekshyyan, OY, Rhoads, CA, Fuseler, J, Harrison, L and Aw, TY (2003) Differential effects of superoxide dismutase isoform expression on hydroperoxide-induced apoptosis in PC-12 cells. J. Biol. Chem. 278:13294-301 |
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| Abstract |
The current study examines the contribution of mitochondria-derived reactive oxygen species (ROS) in tert-butyl-hydroperoxide (TBH)-induced apoptotic signaling using clones of undifferentiated pheochromocytoma (PC-12) cells that stably overexpress the human mitochondrial or cytoplasmic forms of superoxide dismutase (SOD) (viz. Mn-SOD or CuZn-SOD, respectively). Exposure of wild type cells to TBH caused an early generation of ROS (30 min) that resulted in cell apoptosis at 24 h. These responses were attenuated with N-acetylcysteine pretreatment; however, N-acetylcysteine was ineffective in cytoprotection when added after TBH-induced ROS formation. Stable overexpression of SOD isoforms caused a 2- and 3.5-fold elevation in CuZn-SOD and Mn-SOD activities in the cytoplasm and mitochondria, respectively, and 3-fold increases in cellular GSH content. Accordingly, the stable overexpression of Mn-SOD attenuated TBH-induced mitochondrial ROS generation and cell apoptosis. Whereas transient Mn-SOD expression similarly prevented PC-12 apoptosis, this was associated with increases in SOD activity but not GSH, indicating that cytoprotection by Mn-SOD overexpression is related to mitochondrial ROS elimination and not due to increases in cellular GSH content per se. Stable or transient CuZn-SOD overexpression exacerbated cell apoptosis in conjunction with accelerated caspase-3 activation, regardless of cell GSH levels. Collectively, our results support a role for mitochondrial ROS in TBH-induced PC-12 apoptosis that is attenuated by Mn-SOD overexpression and is independent of cellular GSH levels per se. |
| Links |
PubMed Online version:10.1074/jbc.M208670200 |
| Keywords |
Animals; Apoptosis/drug effects; Apoptosis/physiology; Caspase 3; Caspases/metabolism; Enzyme Activation/drug effects; Glutathione/metabolism; Glutathione Disulfide/metabolism; Humans; Isoenzymes/genetics; Isoenzymes/metabolism; PC12 Cells; Rats; Reactive Oxygen Species/metabolism; Superoxide Dismutase/genetics; Superoxide Dismutase/metabolism; Transfection; tert-Butylhydroperoxide/pharmacology |
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