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。
DNA double strand breaks （DSBs） are the most serious DNA damage types in the cell. DSBs can activate cell＇s DNA damage response （DDR） mechanism and then the histone H2AX be rapidly phosphorylated （phosphorylated histone H2AX is called the γ-H2AX）. Subsequently, γ-H2AX gathered in double-stranded breaks and formed a large number of γ-H2AX foci. Therefore, γ-H2AX loci tested in the cell can be used for evaluation of double-stranded DNA rupture, which can be used to evaluate the genetic toxicity of the mutagenic capability. At home and abroad at present, there are many detection methods of γ-H2AX foci, including Western blotting, immunofluorescence staining microscopic method, whole-cell enzyme-linked immune assay and flow cytometry as- say. Each method has its advantages and disadvantages. This paper will be focused on new progress in γ-H2AX detection methods in recent years at home and abroad, and summarize the operation steps, statistical parameters, advantages and disadvantages between different test methods, and analyze the application of γ-H2AX assay in the field of genetic toxicology.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2016年第4期418-424,共7页
Chinese Journal of New Drugs
基金
上海市科技人才项目基金(15XD1523900)
