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30 September 2011 Comprehensive Profiling of Radiosensitive Human Cell Lines with DNA Damage Response Assays Identifies the Neutral Comet Assay as a Potential Surrogate for Clonogenic Survival
Shareef A Nahas, Robert Davies, Francesca Fike, Kotoka Nakamura, Liutao Du, Refik Kayali, Nathan T Martin, Patrick Concannon, Richard A Gatti
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Abstract

In an effort to explore the possible causes of human radiosensitivity and identify more rapid assays for cellular radiosensitivity, we interrogated a set of assays that evaluate cellular functions involved in recognition and repair of DNA double-strand breaks: (1) neutral comet assay, (2) radiation-induced γ-H2AX focus formation, (3) the temporal kinetics of structural maintenance of chromosomes 1 phosphorylation, (4) intra-S-phase checkpoint integrity, and (5) mitochondrial respiration. We characterized a unique panel of 19 “radiosensitive” human lymphoblastoid cell lines from individuals with undiagnosed diseases suggestive of a DNA repair disorder. Radiosensitivity was defined by reduced cellular survival using a clonogenic survival assay. Each assay identified cell lines with defects in DNA damage response functions. The highest concordance rate observed, 89% (17/19), was between an abnormal neutral comet assay and reduced survival by the colony survival assay. Our data also suggested that the neutral comet assay would be a more rapid surrogate for analyzing DNA repair/processing disorders.

Shareef A Nahas, Robert Davies, Francesca Fike, Kotoka Nakamura, Liutao Du, Refik Kayali, Nathan T Martin, Patrick Concannon, and Richard A Gatti "Comprehensive Profiling of Radiosensitive Human Cell Lines with DNA Damage Response Assays Identifies the Neutral Comet Assay as a Potential Surrogate for Clonogenic Survival," Radiation Research 177(2), 176-186, (30 September 2011). https://doi.org/10.1667/RR2580.1
Received: 31 January 2011; Accepted: 1 August 2011; Published: 30 September 2011
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