Histone H2AX phosphorylation as a molecular pharmacological marker for DNA interstrand crosslink cancer chemotherapy
Histone H2AX phosphorylation as a molecular pharmacological marker for DNA interstrand crosslink cancer chemotherapy
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DOI：
10.1016/j.bcp.2008.03.025
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发表时间：
2008-07-01
影响因子：
5.8
通讯作者：
Hartley, J. A.
中科院分区：
文献类型：
--
作者：
Clingen, P. H.;Wu, J. Y. -H.;Hartley, J. A.
本研究的目的是研究DNA链间交联（ICL）剂参与H2 AX磷酸化的作用机制，并确定γ H2 AX是否可以作为鉴定潜在ICL细胞化学敏感性的合适药理学标记。在正常人成纤维细胞中，在用氮芥（HN 2）或顺铂处理后，在使用彗星试验（一种用于检测体外或临床样品中的ICL的经验证的方法）测量的DNA ICL的峰值后2-3小时检测到峰值γ H2 AX响应。与使用彗星试验检测ICL相比，通过免疫荧光显微镜检测γ H2 AX病灶可以常规检测到6-10倍低浓度的两种药物。用于修复DNA ICL的主要途径是通过ERCC 1-XPF内切核酸酶初始脱钩ICL，随后同源重组。HN 2或顺铂诱导的γ H2 AX病灶在ERCC 1或XRCC 3（同源重组）缺陷型中国仓鼠细胞中持续时间显著更长，与野生型或电离辐射敏感的XRCC 5细胞相比，ERCC 1或XRCC 3（同源重组）缺陷型中国仓鼠细胞对ICL试剂的细胞杀伤高度敏感。使用γ H2 AX免疫荧光优于彗星试验的优点是，它似乎可以检测ERCC 1和HR缺陷细胞中的ICL化学敏感性。与化学抗性（A2780 cisR）细胞相比，在HN 2和顺铂的情况下，γ H2 AX病灶也持续存在于化学敏感的人卵巢癌细胞（A2780）中。这些结果表明，γ H2 AX可以作为由HN 2或顺铂诱导的DNA损伤的高度敏感和一般的标志物，并显示出预测潜在的细胞对ICL试剂的化学敏感性的希望。(c)2008年爱思唯尔公司All rights reserved.
The aims of this study were to investigate mechanisms of action involved in H2AX phosphorylation by DNA interstrand crosslinking (ICL) agents and determine whether gamma H2AX could be a suitable pharmacological marker for identifying potential ICL cellular chemosensitivity. In normal human fibroblasts, after treatment with nitrogen mustard (HN2) or cisplatin, the peak gamma H2AX response was detected 2-3 h after the peak of DNA ICLs measured using the comet assay, a validated method for detecting ICLs in vitro or in clinical samples. Detection of gamma H2AX foci by immunofluorescence microscopy could be routinely detected with 6-10 times lower concentrations of both drugs compared to detection of ICLs using the comet assay. A major pathway for repairing DNA ICLs is the initial unhooking of the ICL by the ERCC1-XPF endonuclease followed by homologous recombination. HN2 or cisplatin-induced gamma H2AX foci persisted significantly longer in both, ERCC1 or XRCC3 (homologous recombination) defective Chinese hamster cells that are highly sensitive to cell killing by ICL agents compared to wild type or ionising radiation sensitive XRCC5 cells. An advantage of using gamma H2AX immunofluorescence over the comet assay is that it appears to detect ICL chemosensitivity in both ERCC1 and HR defective cells. With HN2 and cisplatin, gamma H2AX foci also persisted in chemosensitive human ovarian cancer cells (A2780) compared to chemoresistant (A2780cisR) cells. These results show that gamma H2AX can act as a highly sensitive and general marker of DNA damage induced by HN2 or cisplatin and shows promise for predicting potential cellular chemosensitivity to ICL agents. (c) 2008 Elsevier Inc. All rights reserved.