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Phosphorylated fraction of H2AX as a measurement for DNA damage in cancer cells and potential applications of a novel assay

  1. Author:
    Ji, Jay
    Zhang, Yiping
    Redon, Christophe E.
    Reinhold, William C.
    Chen, Alice P.
    Fogli, Laura K.
    Holbeck, Susan L.
    Parchment, Ralph
    Hollingshead, Melinda
    Tomaszewski, Joseph E.
    Dudon, Quentin
    Pommier, Yves
    Doroshow, James H.
    Bonner, William M.

  2. Author Address

    Leidos Biomed Res Inc, Frederick Natl Lab Canc Res, Appl Dev Res Directorate, Frederick, MD 21701 USA.NCI, Dev Therapeut Branch, Ctr Canc Res, Bethesda, MD 20892 USA.NCI, Early Clin Trials Dev Program, Div Canc Treatment & Diag, Bethesda, MD 20892 USA.NCI, Div Canc Treatment & Diag, Bethesda, MD 20892 USA.
    1. Year: 2017
    2. Date: Feb 3
  2. Journal: PLOS ONE
  3. PUBLIC LIBRARY SCIENCE,
    1. 12
    2. 2
    3. Pages: e0171582
  5. Type of Article: Article
  6. Article Number: e0171582
  7. ISSN: 1932-6203
  1. Abstract:

    Phosphorylated H2AX (gamma-H2AX) is a sensitive marker for DNA double-strand breaks (DSBs), but the variability of H2AX expression in different cell and tissue types makes it difficult to interpret the meaning of the gamma-H2AX level. Furthermore, the assays commonly used for gamma-H2AX detection utilize laborious and low-throughput microscopy-based methods. We describe here an ELISA assay that measures both phosphorylated H2AX and total H2AX absolute amounts to determine the percentage of gamma-H2AX, providing a normalized value representative of the amount of DNA damage. We demonstrate the utility of the assay to measure DSBs introduced by either ionizing radiation or DNA-damaging agents in cultured cells and in xenograft models. Furthermore, utilizing the NCI-60 cancer cell line panel, we show a correlation between the basal fraction of gamma-H2AX and cellular mutation levels. This additional application highlights the ability of the assay to measure gamma-H2AX levels in many extracts at once, making it possible to correlate findings with other cellular characteristics. Overall, the gamma-H2AX ELISA represents a novel approach to quantifying DNA damage, which may lead to a better understanding of mutagenic pathways in cancer and provide a useful biomarker for monitoring the effectiveness of DNA-damaging anticancer agents.

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External Sources

  1. View Full Text
  2. DOI: 10.1371/journal.pone.0171582
  3. PMID: 28158293
  4. PMCID: PMC5291513
  5. View record in Web of Science®
  6. WOS: 000396161700102

Library Notes

  1. Fiscal Year: FY2016-2017
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