An optimized method for detecting gamma-H2AX in blood cells reveals a significant interindividual variation in the gamma-H2AX response among humans

被引：146
作者：
Ismail, Ismail Hassan [1 ]
Wadhra, Tabasum Imran [1 ]
Hammarsten, Ola [1 ]
机构：
[1] Univ Gothenburg, Sahlgrens Univ Hosp, Dept Clin Chem & Transfus Med, SE-41345 Gothenburg, Sweden
来源：
NUCLEIC ACIDS RESEARCH | 2007年 / 35卷 / 05期
关键词：
D O I：
10.1093/nar/gkl1169
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Phosphorylation of histone H2AX on serine 139 (gamma-H2AX, gamma H2AX) occurs at sites flanking DNA double-strand breaks (DSBs) and can provide a measure of the number of DSBs within a cell. Here we describe a rapid and simple flowcytometry-based method, optimized to measure gamma-H2AX in non-fixed peripheral blood cells. No DSB induced signal was observed in H2AX-/-cells indicating that our FACS method specifically recognized gamma-H2AX accumulation. The gamma-H2AX assay was capable of detecting DNA damage at levels 100-fold below the detection limit of the alkaline comet assay. The gamma-H2AX signal was quantitative with a linear increase of the gamma-H2AX signal over two orders of magnitude. We found that all nucleated blood cell types examined, including the short-lived neutrophils induce gamma-H2AX in response to DSBs. Interindividual difference in the gamma-H2AX signal in response to ionizing radiation and the DSB-inducing drug calicheamicin was almost 2-fold in blood cells from patients, indicating that the amount of gamma-H2AX produced in response to a given dose of radiation varies significantly in the human population. This simple method could be used to monitor response to radiation or DNA-damaging drugs.
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页数：10
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