Skip to main content
SpringerLink
Log in

Oxidative stress induces apoptosis in C6 glioma cells: Involvement of mitogen-activated protein kinases and nuclear factor kappa B

  • Published:
Neurotoxicity Research Aims and scope Submit manuscript

Abstract

Excessive oxidative stress has been implicated in the induction of cell death in a variety of neurodegenerative diseases. In the present study, hydrogen peroxide (H2O2)-induced cell death in rat C6 glioma cells was used as a model system for studying the molecular events associated with oxidative stress-induced cell death in glial cells. We demonstrate that exposure of C6 glioma cells to H2O2 results in apoptotic cell death in a concentration-dependent manner, and caused activation of a member of the caspase-3-like family of proteases resulting in cleavage of the DNA repair enzyme poly(ADP-ribose)polymerase, PARP. Furthermore, H2O2 induced a transient activation of the transcription factor, nuclear factor kappa B (NFkB). Pre-treatment of cells with the antioxidant N-acetyl-cysteine, (NAC), prevented both the activation of NFkB and the induction of apoptosis by H2O2, suggesting a possible role for this transcription factor in oxidant-induced apoptosis in glial cells. Exposure of the cells to H2O2 led to transient activation of both c-JunN-terminal kinase (JNK) and p38 kinase but has no effect on extracellular regulated kinase (ERK) activity. Inhibition of p38 by SB203580 did not protect the cells against H2O2-induced apoptosis suggesting that activation of p38 is not essential for H2O2-mediated cell death in C6 glioma cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson, P. (1997) Kinase cascades regulating entry into apoptosis.Microbiol. Mol. Biol. Rev. 61, 33–46.

    PubMed  CAS  Google Scholar 

  • Baeuerle, P.A., Henkel, T. (1994) Function and activation of NFkB in the immune system.Annu Rev Immunol 12, 141–179.

    PubMed  CAS  Google Scholar 

  • Barinaga, M. (1998a) Is apoptosis key in alzheimer’s disease?Science 281, 1303–1304.

    Article  PubMed  CAS  Google Scholar 

  • Barinaga, M. (1998b) Stroke-damaged neurons may commit cellular suicide.Science 281, 1302–1303.

    Article  PubMed  CAS  Google Scholar 

  • Beg, A.A. and Baltimore, D. (1996) An essential role for NFkB in preventing TNF-alpha-induced cell death.Science 274, 782–784.

    Article  PubMed  CAS  Google Scholar 

  • Bhat, R.B. and Zhang, P. (1999) Hydrogen peroxide activation of multiple mitogen-activated protein kinases in an oligodendrocyte cell line: role of extracellular signal-regulated kinase in hydrogen peroxide-induced cell death.J. Neurochem. 72, 112–119.

    Article  PubMed  CAS  Google Scholar 

  • Boland, M.P., Foster, S.J. and O’Neill, L.A.J. (1997) Daunoru-bicin activates NFkB and induce KB-dependent gene expression in HL-60 promyelocytic and Jurkat T lymphoma cells.J. Biol. Chem. 272, 12952–12960.

    Article  PubMed  CAS  Google Scholar 

  • Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principal of protein-dye binding.Anal. Biochem. 73, 248–254.

    Article  Google Scholar 

  • Casano, F.J., Rolando, A.M., Mudgett, J.S. and Molineaux, S.M. (1994) The structure and complete nucleotide sequence of the murine gene encoding interleukin-1β converting enzyme (ICE).Genomics 20, 474–481.

    Article  PubMed  CAS  Google Scholar 

  • Chu, Z.L., McKinsey, T.A., Liu, L., Gentry, J.J., Malim, M.H. and Ballard, D.W. (1997) Suppression of tumour necrosis factor-induced cell death by inhibitor of apoptosis c-IAP-2 is under NFkappaB control.Proc. Natl. Acad. Sci. U.S.A. 94, 10057–10063.

    Article  PubMed  CAS  Google Scholar 

  • Cohen, G.M. (1997) Caspases: the executioners of apoptosis.Biochem. J. 326, 1–16.

    PubMed  CAS  Google Scholar 

  • Cowden, R.R. and Curtis, S.K. (1981) Microfluorimetric investigations of chromatin structure.Histochemistry 72, 11–23.

    Article  PubMed  CAS  Google Scholar 

  • Gardner, A.M. and Johnson, G.L. (1996) Fibroblast growth factor-2 suppression of tumor necrosis factor-mediated apoptosis requires ras and the activation of mitogen-activated protein kinase.J. Biol. Chem. 271, 14560–14566.

    Article  PubMed  CAS  Google Scholar 

  • Giri, D.K. and Aggarwal, B.B. (1998) Constitutive activation of NFkB causes resistance to apoptosis in human cancerous T cell lymphoma HuT 78 cells.J. Biol. Chem. 273, 14008–14014.

    Article  PubMed  CAS  Google Scholar 

  • Gupta, S., Campbell, D., Derijard, B. and Davis, R.J. (1995) Transcription factor ATF2 regulation by the JNK signal transduction pathway.Science 267, 389–393.

    Article  PubMed  CAS  Google Scholar 

  • Gwag, B.J., Lobner, D., Koh, J. and Choi, D.W. (1994) Blockade of glutamate receptors during oxygen or glucose deprivation unmasks apoptosis in cultured cortical neurons.Neurosci. Abs. 15, 248.

    Google Scholar 

  • Hampton, M.B. and Orrenius, S. (1997) Dual regulation of caspase activity by hydrogen peroxide: implication for apoptosis.FEBS lett. 414, 552–556.

    Article  PubMed  CAS  Google Scholar 

  • Janicke, R.U., Ng, P., Sprengart, ML., Porter, A.G. (1998) Caspase-3 is required for alpha-fodrin cleavage but dispensable for cleavage of other death substrates in apoptosis.J. Biol. Chem. 273, 15540–15545.

    Article  PubMed  CAS  Google Scholar 

  • Kitamura, Y., Ota, T., Matsuoka, Y., Tooyama, I., Kimura, H., Shimohama, S., Nomura, Y., Gebicke-Haerter, P.J. and Taniguchi, T. (1999) Hydrogen peroxide-induced apoptosis mediated by p53 protein in glial cells.Glia 25, 154–164.

    Article  PubMed  CAS  Google Scholar 

  • Liu, X.Z., Xu, X.M., Hu, R., Du, C, Zhang, S.X., McDonald, J.W., Dong, H.X., Wu, Y.J., Fan, G.S., Jacquin, M.F., Hsu, C.Y., Choi, D.W. (1997) Neuronal and glial apoptosis after traumatic spinal cord injury.J. Neurosci. 17, 5395–5406.

    PubMed  CAS  Google Scholar 

  • Lassman, H., Bancher, C, Breitschopf, H., Wiegel, J., Bobin-ski, M., Jellinger, K. and Wisniewski, H.M. (1995) Cell death in Alzheimer’s disease evaluated by DNA fragmentation in situ.Acta. Neuropathol. 89, 35–41.

    Article  Google Scholar 

  • Martin, S.J., Newmeyer, D.D., Mathias, S., Farschon, D.M., Wang, H., Reed, J.C. and Green, D.R. (1995) Cell free reconstitution of Fas-, UV radiation- and cera-mide-induced apoptosis.EMBO J. 14, 5191–5200.

    PubMed  CAS  Google Scholar 

  • Meyer, M., Pahl, H.L. and Baeuerle, P.A. (1994) Regulation of the transcription factors NF-kappa B and AP-1 by redox changes.Chem. Biol. Interact. 91, 91–100.

    Article  PubMed  CAS  Google Scholar 

  • Mosieniak, G., Figiel, I. and Kaminska, B. (1997) Cyclosporin A, an immunosuppressive drug induces programmed cell death in rat C6 glioma cells by a mechanism that involves the AP-1 transcription factor.J. Neurochem. 68, 1142–1149.

    PubMed  CAS  Google Scholar 

  • Moynagh, P.N., Williams, D.C. and O’Neill, L.A.J. (1993) Interleukin-1 activates transcription factor NF-kappa-B in glial cells.Biochem. J. 294, 343–347.

    PubMed  CAS  Google Scholar 

  • O’Connell, M.A., Cleere, R., Long, A., O’Neill, L.A.J and Kel-leher D. (1995) Cellular proliferation and activation of NF-kappa B are induced by autocrine production of tumour necrosis factor alpha in the human T lymphoma line HuT 78.J. Biol. Chem. 270, 7399–7404.

    Article  PubMed  CAS  Google Scholar 

  • O’Neill, L.A.J and Kaltschmidt, C. (1997) NFkB: a crucial transcription factor for glial and neuronal cell function. Trends Neuro Sci20: 252–258.

    Article  CAS  Google Scholar 

  • Ozaki, I., Tani, E., Ikemoto, H., Kitagawa, H. and Fujikawa, H. (1999) Activation of stress-activated protein kinase/c-Jun NH2-terminal kinase and p38 kinase in cal-phostin C-induced apoptosis requires caspase-3-like proteases but is indispensable for cell death. J. Biol. Chem.274, 5310–5217.

    Article  PubMed  CAS  Google Scholar 

  • Polverino, A.J. and Patterson, S.D. (1997) Selective activation of caspases during apoptotic induction of HL-60 cells.J. Biol. Chem. 272, 7013–7021.

    Article  PubMed  CAS  Google Scholar 

  • Robinson, M.J. and Cobb, M.H. (1997) Mitogen-activated protein kinase pathways.Curr. Opin. Cell Biol. 9, 180186.

    Article  Google Scholar 

  • Schulze-Osthoff, K., Krammer, P.H. and Droge W. (1994) Divergent signalling via APO-1/Fas and the TNF recepor, two homologous molecules involved in physiological cell death.EMBO J. 13, 4587–4596.

    PubMed  CAS  Google Scholar 

  • Schraufstatter, I.U., Hyslop, P.A., Hinshaw, D.B., Spragg, R.G., Sklar, L..A and Cochrane, C.G. (1986) Hydrogen peroxide induced injury of cells and its prevention by inhibitors of poly(ADP-ribose)polymerase.Proc. Natl. Acad. Sci. USA. 83, 4908–4912.

    Article  PubMed  CAS  Google Scholar 

  • Siman, R., Bozyczko-Coyne, D., Meyer, S.L. and Bhat RV. (1999) Immunolocalization of caspase proteolysis in situ: evidence for widespread caspase-mediated apoptosis of neurons and glia in the postnatal rat brain.Neuroscience 92, 1425–1442.

    Article  PubMed  CAS  Google Scholar 

  • Simonian, N.A., Coyle, J.T. (1996) Oxidative stress in neurodegenerative diseases. Annu. Rev. Pharmacol. Toxicol.36, 83–106.

    Article  PubMed  CAS  Google Scholar 

  • Smale G., Nichols NR, Brady DR, Finch CE, Horton WE. (1995) Evidence for apoptotic cell death in Alzheimer’s disease.Exp. Neurol. 133, 225–230.

    Article  PubMed  CAS  Google Scholar 

  • Stridh, H., Kimland, M., Jones, D.P., Orrenius, S., Hampton, M.B. (1998) Cytochrome C release and caspase activation in hydrogen peroxide- and tributyltin-induced apoptosisFEBS lett. 429, 351–355.

    Article  PubMed  CAS  Google Scholar 

  • Thomas, E.L., Grisham, M.B., Melton, D.F. and Jefferson, M.M. (1985) Evidence for a role of taurine in the in vitro oxidative toxicity of neutrophils towards erythrocytes.J. Biol. Chem. 260, 3321–3329.

    PubMed  CAS  Google Scholar 

  • Thornberry, N.A., Rano, T.A., Peterson, E.P., Rasper, D.M., Timkey, T., Garcia-Calvo, M., Houtzager, V.M., Nord-strum, P.A., Roy, S., Vaillancourt, J.P., Chapman, K.T. and Nicholson, D.W. (1997) A combinatorial approach defines specificities of members of the caspase family and granzyme B.J. Biol. Chem. 272, 17907–17911.

    Article  PubMed  CAS  Google Scholar 

  • Whittemore, E.R., Loo, D.T., Cotman C.W. (1994) Exposure to hydrogen peroxide induces cell death via apoptosis in cultured rat cortical neurons.Neuroreport 5, 1485–1488.

    Article  PubMed  CAS  Google Scholar 

  • Wu, M. and Lozano, G. (1994) NFkB activation of p53. A potential mechanism for suppressing cell growth in response to stress.J. Biol. Chem. 269, 20067–20074.

    CAS  Google Scholar 

  • Wylie, A.H., Kerr, J. FR. and Currie, A.R. (1980) Cell death: the significance of apoptosis.Int. Rev. Cytol. 68, 251–306.

    Article  Google Scholar 

  • Xiang, J., Chao, D.T. and Korsmeyer, S.J. (1996) Bax-induced cell death may not require interleukin-1-converting enzyme-like proteases.Proc. Natl. Acad. Sci. U.S.A. 93, 14559–14563.

    Article  PubMed  CAS  Google Scholar 

  • Yasugi, E., Nakata, K., Yokoyama, Y., Kano, K., Dohi, T. and Oshima, M. (1998) Dihydroheptoprenyl and dihydro-decprenyl monophosphate induce apoptosis mediated by activation of caspase-3-like protease.Biochim. Biophys. Acta. 1389, 132–140.

    PubMed  CAS  Google Scholar 

  • Yoshimura, S-I., Sakai, H., Ohguchi, K., Nakashima, S., Banno, Y., Nishimura, Y., Sakai, N., Nozawa, Y. (1997) Changes in the activity and mRNA levels of phospholi-pase D during ceramide-induced apoptosis in rat C6 glial cells.J. Neurochem. 69, 713–720.

    Article  PubMed  CAS  Google Scholar 

  • Zisterer, D.M., Hance, H., Campiani, G., Garofalo, A., Nacci, V. and Williams, D.C. (1998) Antiproliferative action of pyrrolobenzoxazepine derivatives in cultured cells.Biochem. Pharmacol. 55, 397–403.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Marina Marangolo

    Present address: Westmead Institute for Cancer Research, Westmead Millennium Institute, Darcy Road, 2145, Westmead, NSW, Australia

Authors and Affiliations

  1. Department of Biochemistry, Trinity College Dublin, Dublin 2, Ireland

    Marina Marangolo, Margaret M. McGee, Keith F. Tipton, D. Clive Williams & Daniela M. Zisterer

Authors
  1. Marina Marangolo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Margaret M. McGee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Keith F. Tipton
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Clive Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daniela M. Zisterer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Margaret M. McGee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marangolo, M., McGee, M.M., Tipton, K.F. et al. Oxidative stress induces apoptosis in C6 glioma cells: Involvement of mitogen-activated protein kinases and nuclear factor kappa B. neurotox res 3, 397–409 (2001). https://doi.org/10.1007/BF03033200

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03033200

Keywords

Search

Navigation