Measurement of DNA Interstrand Crosslinking in Individual Cells Using the Single Cell Gel Electrophoresis (Comet) Assay
Measurement of DNA Interstrand Crosslinking in Individual Cells Using the Single Cell Gel Electrophoresis (Comet) Assay
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DOI：
10.1007/978-1-60327-418-0_17
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Hartley, John A.
中科院分区：
文献类型：
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作者：
Spanswick, Victoria J.;Hartley, Janet M.;Hartley, John A.
单细胞凝胶电泳(彗星)试验最初是为了显示单个细胞中的DNA链断裂损伤，现在已经被用于测量DNA链间的交联链。DNA链间交叉连接是由一些常用的癌症化疗药物在细胞内形成的，被认为是此类药物形成的关键损伤。这项技术可以在单个细胞水平上分析DNA链间交联的形成和修复，需要很少的细胞，可以确定细胞群体内反应的异质性，并且足够灵敏，可以在药理上相关的剂量下测量DNA链间交联。该方法可应用于任何可获得单细胞悬浮液的体外或体内应用。该方法在使用人体组织的研究中也变得非常有价值，可以作为早期临床试验中的药效学分析方法。
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 arc 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.