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Acta Biochimica et Biophysica Sinica Advance Access originally published online on July 5, 2009
Acta Biochimica et Biophysica Sinica 2009 41(8):625-630; doi:10.1093/abbs/gmp050
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© The Author 2009. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

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

Wenli Yuan1,2, Jiazhi Guo1, Xingguo Li1, Zhirong Zou1, Guangxue Chen3, Jun Sun1, Tinghua Wang3,* and Di Lu1,*

1 Department of Anatomy, Kunming Medical University, Kunming 650031, China
2 Department of Clinical Laboratory, Second Hospital of Yunnan Province, Kunming 650021, China
3 Institute for Research on Neuroscience, Kunming Medical University, Kunming 650031, China

* Correspondence address. Tel/Fax: +86-871-5342766; E-mail: tinghua_neuron{at}263.net (T.W.); Tel/Fax: +86-871-5316881; E-mail: ludi20040609{at}126.com (D.L.)


   Abstract

It has been reported that phospholipase C-{gamma}1 (PLC-{gamma}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-{gamma}1 activation remain to be identified. The present study was designed to examine the roles of PLC-{gamma}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-{gamma}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 [GenBank] , 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 [GenBank] , Bcl-2 protein expression was dramatically decreased and the activation of pro-caspase-3 was significantly increased. We concluded that PLC-{gamma}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    phospholipase C-{gamma}1; hydrogen peroxide; apoptosis; Bcl-2; caspase-3

Received: January 15, 2009; Accepted: April 28, 2009
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