生物学杂志 ›› 2020, Vol. 37 ›› Issue (1): 16-.doi: 10.3969/j.issn.2095-1736.2020.01.016

• 研究报告 • 上一篇    下一篇

γH2AX、53BP1及RAD51焦点用于分析DNA双链断裂损伤

  

  1. 1. 山西医科大学 第二临床医学院, 太原 030013; 2. 山西医科大学附属肿瘤医院研究所, 太原 030013
  • 出版日期:2020-02-18 发布日期:2020-03-09
  • 通讯作者: 苏文,博士,硕士生导师,主要研究方向为肿瘤免疫学,E-mail:suwen30@yahoo.com
  • 作者简介:王依朝,硕士研究生,研究方向为肿瘤学,E-mail:wangyz270@163.com
  • 基金资助:
    国家自然科学基金(批准号:81272696);山西省重大研发计划(社会发展)项目 (批准号: 201703D321013)

γH2AX, 53BP1 and RAD51 foci analysis for monitoring DNA double-strand breaks

  1. 1. The Second Clinical Medical College, Shanxi Medical University, Taiyuan 030013;
     2. Research Institute of Tumor Hospital Affiliated to Shanxi Medical University, Taiyuan 030013, China
  • Online:2020-02-18 Published:2020-03-09

摘要: γH2AX、53BP1及RAD51焦点(foci)是DNA双链断裂(DSBs)损伤修复的评价指标。其中,γH2AX分析应用最为广泛,但最新有研究发现其并不完全与DSBs损伤一致。系统分析了DNA损伤时γH2AX、53BP1及RAD51 foci的动力性变化及其在DSBs损伤评估中的合理应用。结果表明,在电离辐射处理时,γH2AX和53BP1 foci具有高度一致性,均可反映DSBs损伤及修复的动力性变化,而RAD51 foci仅出现在部分损伤细胞中,与其主要参与同源重组修复一致。在喜树碱(CPT)处理时,γH2AX呈现两种染色类型(亮而离散的foci和泛核磷酸化),53BP1 foci仅出现在亮而离散的γH2AX foci阳性细胞且与其高度一致。结果提示,单独γH2AX染色阳性并不能准确代表DSBs损伤,而应结合53BP1 等染色来综合分析DSBs损伤程度;RAD51 foci分析仅能反映部分细胞的DSBs损伤修复(主要是同源重组)情况。

关键词: DNA双链断裂, DNA修复, 电离辐射, 磷酸化的组蛋白H2AX

Abstract: γH2AX, 53BP1 and RAD51 foci are the evaluation markers of DNA double strand breaks (DSBs). Above all, γH2AX foci analysis is the most widely used, but recent studies have found that it is not completely consistent with DSBs damage. This study systematically analyzed the dynamic changes of γH2AX, 53BP1 and RAD51 foci during DNA damage and their reasonable application in DSBs damage assessment. According to the results, γH2AX and 53BP1 foci were highly consistent after cells exposed to ionizing radiation treatment, they both could reflect accurately the dynamic changes of DSBs damages and repairs; while RAD51 foci only appeared in part of damaged cells, which was consistent with its role mainly involved in homologous recombination process. Upon treatment with Camptothecin (CPT), γH2AX presented two types of staining (bright and discrete foci and pan-nuclear phosphorylation), while 53BP1 foci only appeared in cells with bright and discrete γH2AX foci and were highly consistent with γH2AX foci. These results revealed that positive staining of γH2AX alone cannot accurately represent DSBs damage, and the extent of DSBs damage should be comprehensively analyzed by combining with staining of 53BP1. The RAD51 foci analysis only reflect the DSBs damage repair of part cells, mainly homologous recombination repair. 

Key words: DNA double-stand breaks, DNA damage repair, ionize radiation, γH2AX

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