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The Generation of Stable Oxidative Stress-Resistant Phenotypes in Chinese Hamster Fibroblasts Chronically Exposed to Hydrogen Peroxide or Hyperoxia

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Free Radicals and Antioxidant Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 610))

Abstract

With the recognition that chronic exposure to oxidative stress occurs in many disease states and can be deleterious to the human health, great interest has emerged in understanding the mechanisms by which mammalian cells develop resistance to chronic oxidative stress. In order to study the mechanisms of development of resistance to chronic oxidative stress, a model system where Chinese hamster fibroblasts (HA1) are chronically exposed to progressively increasing concentrations of H2O2 (50–800 μM) or hyperoxia (80–95% O2) has been developed. Following >200 d of exposure to H2O2 (or 18 months of exposure to hyperoxia), the cells developed stable H2O2-resistant (or O2-resistant) phenotypes that are characterized by increases in total glutathione, antioxidant enzyme activity, heme oxygenase activity, stress protein gene expression, DNA repair pathways, and resistance to a wide variety of other toxic stress known to cause oxidant injury. In addition, these oxidant-resistant cells exhibited amplification of the gene for catalase and constitutively elevated AP-1 DNA binding activity. Further, beyond 240 d genomic instability as evidenced by chromosomal rearrangements and alterations in ploidy was stably maintained upon removal from the chronic oxidative stress conditions. These results demonstrate the capability of mammalian cells to develop stable oxidative stress-resistant phenotypes in response to both exogenous (H2O2) as well as endogenous (95% O2) oxidative stress. The understanding of mechanisms of resistance to oxidative stress and its possible relevance in various disease states are discussed.

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Acknowledgments

DRS is supported by NIH R01-CA100045, DOE DE-FG02-02ER63447, and NIEHS P42 ES013661. SJS is supported by the Department of Pediatrics at the University of Iowa.

Author information

Authors and Affiliations

  1. Departments of Radiation Oncology, Holden Comprehensive Cancer Center, The University of Iowa, Iowa City, IA, USA

    Douglas R. Spitz

  2. Department of Pediatrics, The University of Iowa, Iowa City, IA, USA

    Shannon J. Sullivan

Authors
  1. Douglas R. Spitz
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  2. Shannon J. Sullivan
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Editor information

Editors and Affiliations

  1. A& M College, Southern University &, James L. Hunt Street, Baton Rouge, 70813, U.S.A.

    Rao M. Uppu

  2. School of Medicine, Tulane University, Tulane Ave. 1430, New Orleans, 70112, U.S.A.

    Subramanyam N. Murthy

  3. Biodynamics Inst., Louisiana State University, Baton Rouge, 70808, U.S.A.

    William A. Pryor

  4. Dorothy M. Davis Heart &, Ohio State University, West 12th Ave. 473, Columbus, 43210, U.S.A.

    Narasimham L. Parinandi

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Spitz, D.R., Sullivan, S.J. (2010). The Generation of Stable Oxidative Stress-Resistant Phenotypes in Chinese Hamster Fibroblasts Chronically Exposed to Hydrogen Peroxide or Hyperoxia. In: Uppu, R., Murthy, S., Pryor, W., Parinandi, N. (eds) Free Radicals and Antioxidant Protocols. Methods in Molecular Biology, vol 610. Humana Press. https://doi.org/10.1007/978-1-60327-029-8_11

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  • DOI: https://doi.org/10.1007/978-1-60327-029-8_11

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-710-5

  • Online ISBN: 978-1-60327-029-8

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