Details

Autor(en) / Beteiligte

• Ji, Jiuping
• Zhang, Yiping
• Redon, Christophe E
• Reinhold, William C
• Chen, Alice P
• Fogli, Laura K
• Holbeck, Susan L
• Parchment, Ralph E
• Hollingshead, Melinda
• Tomaszewski, Joseph E
• Dudon, Quentin
• Pommier, Yves
• Doroshow, James H
• Bonner, William M
• Huen, Michael Shing-Yan

Titel

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

Ist Teil von

• PloS one, 2017-02, Vol.12 (2), p.e0171582

Ort / Verlag

United States: Public Library of Science

Erscheinungsjahr

2017

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• Phosphorylated H2AX (γ-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 γ-H2AX level. Furthermore, the assays commonly used for γ-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 γ-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 γ-H2AX and cellular mutation levels. This additional application highlights the ability of the assay to measure γ-H2AX levels in many extracts at once, making it possible to correlate findings with other cellular characteristics. Overall, the γ-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.