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

Authors: Yuan, Wenli; Guo, Jiazhi; Li, Xingguo; Zou, Zhirong; Chen, Guangxue; Sun, Jun; Wang, Tinghua; Lu, Di

Source: Acta Biochimica et Biophysica Sinica, 5 August 2009, vol. 41, no. 8, pp. 625-630(6)

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Abstract:

It has been reported that phospholipase C-1 (PLC-1) plays an important protective role in hydrogen peroxide (H2O2)-induced pheochromocytoma (PC) 12 cells death. However, most studies have used high doses of H2O2 and the downstream targets of PLC-1 activation remain to be identified. The present study was designed to examine the roles of PLC-1 signaling pathway in the apoptosis of PC12 cells induced by low dose of H2O2, as well as the downstream factors involved in this pathway. Low-dose treatment of H2O2 resulted in PLC-1 tyrosine phosphorylation in a time-dependent manner and H2O2 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 H2O2 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-1 plays an important protective role in H2O2-induced PC12 cells death. Bcl-2 and caspase-3 probably participate in the signaling pathway as downstream factors.

Keywords: Bcl-2; apoptosis; caspase-3; hydrogen peroxide; phospholipase C-1

Document Type: Research Article

DOI: http://dx.doi.org/10.1093/abbs/gmp050

Publication date: August 5, 2009

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