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Roles of Ras-Erk in apoptosis of PC12 cells induced by trophic factor withdrawal or oxidative stress

Journal of Molecular Neuroscience volume 25, pages 133–140 (2005)Cite this article

Abstract

To understand the role of Ras-MAPK (mitogen-activated protein kinase) in trophic factor withdrawal- and oxidative stress-induced apoptotic cell death processes, undifferentiated rat pheochromocytoma PC12 cells and a PC12 variant cell line stably expressing the Ras dominant-negative mutant (M-M17-26) were subjected to serum withdrawal in the absence or presence of H2O2 treatment. The extent of cell death was analyzed by lactate dehydrogenase release, internucleosomal DNA fragmentation, and caspase-3 assays. Both serum with-drawal and H2O2 treatment induced apoptotic cell death in PC12 cells, and the extent of cell death was greatly enhanced in M-M17-26 cells. DNA fragmentation induced by serum withdrawal or H2O2 treatment was blocked completely by a general caspase inhibitor, Z-VAD-FMK. A selective MAPK kinase inhibitor, U0126, blocked the H2O2-induced phosphorylation of Erk1/2 (extracellular signal-regulated kinase) in PC12 cells and increased the levels of active caspase-3 in M-M17-26 under serum withdrawal or H2O2 treatment. In addition, the short-term H2O2 treatment (5–30 min) was sufficient to cause DNA fragmentation in M-M17-26 cells even though H2O2 was removed and cells were incubated in regular growth medium with complete serum for 24 h. However, similar, short-term H2O2 treatment of PC12 cells did not induce DNA fragmentation 24 h later. These results suggest that the Ras-Erk pathway is critical in mediating protection against apoptotic cell death induced by either trophic factor withdrawal or increased oxidative stress.

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Affiliations

  1. William T. Gossett Neurology Laboratories, USA

    Hao Jiang, Lijie Zhang, David Koubi, Jarret Kuo, Laurent Groc, Alba I. Rodriguez, Tangella Jackson Hunter, Stephen Tang & Robert A. Levine

  2. Complementary and Integrative Medicine Research Program, USA

    Hao Jiang, Lijie Zhang, Jarret Kuo, Alba I. Rodriguez, Tangella Jackson Hunter & Robert A. Levine

  3. Department of Surgery, Henry Ford Health System, 48202, Detroit, MI

    Subhash C. Gautam

  4. John D. Dingell Veterans Administration Medical Center, 48201, Detroit, MI

    Hao Jiang & Robert A. Levine

  5. Department of Pharmaceutical Sciences, Wayne State University School of Pharmacy, 48201, Detroit, MI

    Robert A. Levine

  6. Department of Pharmacology and Experimental Therapeutics, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, 91120, Jerusalem, Israel

    Philip Lazarovici

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  1. Hao Jiang
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Jiang, H., Zhang, L., Koubi, D. et al. Roles of Ras-Erk in apoptosis of PC12 cells induced by trophic factor withdrawal or oxidative stress. J Mol Neurosci 25, 133–140 (2005). https://doi.org/10.1385/JMN:25:2:133

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  • DOI: https://doi.org/10.1385/JMN:25:2:133

Index Entries

  • PC12
  • Ras
  • Erk
  • apoptosis
  • oxidative stress
  • trophic factor
  • hydrogen peroxide