基于γ-H2AX生物标志物DNA遗传毒性检测方法的研究进展

发布时间：2019-08-09 13:58

【摘要】：DNA双链断裂(DNA double strand breaks,DSBs)是细胞DNA损伤类型中最严重的一种。DSBs可以激活细胞的DNA损伤应答(DNA damage response,DDR)机制,从而使组蛋白H2AX迅速磷酸化(磷酸化的H2AX组蛋白被称为γ-H2AX)。随后,γ-H2AX聚集在双链断裂处,形成由大量γ-H2AX聚集成的灶点(foci)。检测细胞中的γ-H2AX灶点数目就可以用于评价DNA双链断裂情况,进而可以用来评价遗传毒性因子的致突变能力。目前,国内外有多种对γ-H2AX灶点的检测方法,包括蛋白质印迹法(Western blotting)、免疫荧光染色显微镜检测法、全细胞酶联免疫反应(Whole-cell ELISA)检测法和流式细胞术检测法等,每种方法各有优缺点。本文将重点阐述近几年国内外γ-H2AX检测方法的新进展,并总结对比不同方法之间的操作步骤、统计参数及优缺点,分析γ-H2AX检测方法在遗传毒理方面的应用前景。
[Abstract]:DNA double strand breaks (DNA double strand breaks,DSBs) is one of the most serious types of DNA damage in cells. DSBs can activate the mechanism of DNA damage response to (DNA damage response,DDR, resulting in the rapid phosphorylation of histone H2AX (phosphorylated H2AX histone is called 纬-H2AX). Subsequently, 纬-H2AX aggregates at double strand breaks, forming (foci)., which is formed by a large number of gamma-H2AX aggregates. The detection of the number of 纬-H2AX foci in cells can be used to evaluate the double strand breaks of DNA, and then to evaluate the mutagenicity of genotoxic factors. At present, there are many methods for the detection of gamma-H2AX foci at home and abroad, including Western printing (Western blotting), immunofluorescence staining microscope, whole-cell enzyme-linked immunosorbent assay (Whole-cell ELISA) and flow cytometry. Each method has its own advantages and disadvantages. In this paper, the new progress of gamma-H2AX detection methods at home and abroad in recent years is described, and the operation steps, statistical parameters, advantages and disadvantages of different methods are summarized and compared, and the application prospect of gamma-H2AX detection methods in genetic toxicology is analyzed.
【作者单位】：国家上海新药安全评价研究中心;
【基金】：上海市科技人才项目基金(15XD1523900)
【分类号】：R99

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