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It has been reported that phospholipase C-gamma1 (PLC-gamma1) plays an important protective role in hydrogen peroxide (H(2)O(2))-induced pheochromocytoma (PC) 12 cells death. However, most studies have used high doses of H2O2 and the downstream targets of PLC-gamma1 activation remain to be identified. The present study was designed to examine the roles of PLC-gamma1 signaling pathway in the apoptosis of PC12 cells induced by low dose of H(2)O(2), as well as the downstream factors involved in this pathway. Low-dose treatment of H(2)O(2) resulted in PLC-gamma1 tyrosine phosphorylation in a time-dependent manner and H(2)O(2) killed the PC12 cells by inducing necrosis. In contrast, pretreatment of PC12 cells with U73122, a specific inhibitor of PLC, markedly increased the percentage of dead cells. The mode of cell death was converted to apoptosis as determined by Hoechst/PI nuclear staining and fluorescence microscopy. Western blot analysis demonstrated that the expression of Bcl-2 protein and the activation of pro-caspase-3 were not significantly affected by low dose of H(2)O(2) alone. However, after pretreatment with U73122, Bcl-2 protein expression was dramatically decreased and the activation of pro-caspase-3 was significantly increased. We concluded that PLC-gamma1 plays an important protective role in H(2)O(2)-induced PC12 cells death. Bcl-2 and caspase-3 probably participate in the signaling pathway as downstream factors.

Hydrogen peroxide induces the activation of the phospholipase C-&gamma;1 survival pathway in PC12 cells: protective role in apoptosis

Hydrogen peroxide induces the activation of the phospholipase C-γ1 survival pathway in PC12 cells: protective role in apoptosis