An optimized method for detecting gamma-H2AX in blood cells reveals a significant interindividual variation in the gamma-H2AX response among humans.
An optimized method for detecting gamma-H2AX in blood cells reveals a significant interindividual variation in the gamma-H2AX response among humans.
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DOI：
10.1093/nar/gkl1169
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发表时间：
2007
影响因子：
14.9
通讯作者：
Hammarsten O
中科院分区：
文献类型：
--
作者：
Ismail IH;Wadhra TI;Hammarsten O
组蛋白H2 AX在丝氨酸139（γ-H2 AX，γ H2 AX）上的磷酸化发生在DNA双链断裂（DSB）侧翼的位点处，并且可以提供细胞内DSB数量的量度。在这里，我们描述了一种快速，简单的流式细胞术为基础的方法，优化测量γ-H2 AX在非固定的外周血细胞。在H2 AX-/-细胞中没有观察到DSB诱导的信号，表明我们的FACS方法特异性识别γ-H2 AX积累。γ-H2 AX测定能够检测到低于碱性彗星测定检测限100倍的水平的DNA损伤。γ-H2 AX信号是定量的，γ-H2 AX信号线性增加超过两个数量级。我们发现，所有检查的有核血细胞类型，包括短寿命的中性粒细胞诱导γ-H2 AX响应DSB。γ-H2 AX信号对电离辐射和DSB诱导药物加利车霉素的响应的个体间差异在患者的血细胞中几乎是2倍，这表明响应于给定剂量的辐射产生的γ-H2 AX的量在人群中变化显著。这种简单的方法可用于监测对辐射或DNA损伤药物的反应。
Phosphorylation of histone H2AX on serine 139 (gamma-H2AX, γH2AX) 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 flow-cytometry-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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影响因子：
4.8
作者：
Furuta, T;Takemura, H;Pommier, Y
通讯作者：
Pommier, Y
DOI：
10.1073/pnas.0501895102
发表时间：
2005-06-21
影响因子：
11.1
作者：
Löbrich, M;Rief, N;Uder, M
通讯作者：
Uder, M
影响因子：
56.9
作者：
Celeste, A;Petersen, S;Nussenzweig, A
通讯作者：
Nussenzweig, A
DOI：
10.1016/0360-3016(95)00068-a
发表时间：
1995-07-30
影响因子：
7
作者：
BROCK, WA;TUCKER, SL;PETERS, LJ
通讯作者：
PETERS, LJ
影响因子：
3.1
作者：
Olive, PL;Durand, RE;Trotter, T
通讯作者：
Trotter, T