使用单细胞凝胶电泳（彗星）试验测量单个细胞中的DNA链间交联。

Measurement of DNA interstrand crosslinking in individual cells using the Single Cell Gel Electrophoresis (Comet) assay.

作者信息

Spanswick Victoria J, Hartley Janet M, Hartley John A

机构信息

Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, University College London, Paul O'Gorman Building, London, UK.

出版信息

Methods Mol Biol. 2010;613:267-82. doi: 10.1007/978-1-60327-418-0_17.

DOI:10.1007/978-1-60327-418-0_17

PMID:19997890

Abstract

The Single Cell Gel Electrophoresis (Comet) assay, originally developed to allow visualisation of DNA strand break damage in individual cells, has been adapted to measure DNA interstrand cross-links. DNA interstrand cross-links are formed in cells by a number of commonly used cancer chemotherapy agents and are considered to be the critical lesion formed by such agents. This technique allows the analysis of DNA interstrand cross-link formation and repair at a single cell level, requires few cells, allows the determination of heterogeneity of response within a cell population and is sensitive enough to measure DNA interstrand cross-links at pharmacologically relevant doses. The method can be applied to any in vitro or in vivo application where a single cell suspension can be obtained. The method has also become invaluable in studies using human tissue and can be used as a method for pharmacodynamic analysis in early clinical trials.

摘要

单细胞凝胶电泳（彗星）试验最初用于可视化单个细胞中的DNA链断裂损伤，现已被用于测量DNA链间交联。DNA链间交联由多种常用的癌症化疗药物在细胞中形成，被认为是这些药物形成的关键损伤。该技术能够在单细胞水平分析DNA链间交联的形成和修复，所需细胞数量少，可确定细胞群体内反应的异质性，并且灵敏度足以测量药理相关剂量下的DNA链间交联。该方法可应用于任何能够获得单细胞悬液的体外或体内实验。该方法在使用人体组织的研究中也变得非常重要，可作为早期临床试验中药效学分析的一种方法。