彗星试验：一种检测单个细胞中DNA损伤的灵敏方法。

The comet assay: a sensitive method for detecting DNA damage in individual cells.

作者信息

Liao Wenjuan, McNutt Michael A, Zhu Wei-Guo

机构信息

Department of Biochemistry and Molecular Biology, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Health Science Center, Beijing, China.

出版信息

Methods. 2009 May;48(1):46-53. doi: 10.1016/j.ymeth.2009.02.016. Epub 2009 Mar 6.

DOI:10.1016/j.ymeth.2009.02.016

PMID:19269328

Abstract

The comet assay is a sensitive and rapid method for DNA strand break detection in individual cells, and the year 2009 represents the 25th anniversary of the first description of this methodology in 1984. Over time this method has been improved, but is still not completely standardized, and variations are currently in widespread use with emphasis on applications in research and genetic toxicology. Here we review the principles of the comet assay and cite key studies that have focused on this assay in the past 25 years. In addition, we present an example of how the comet assay was used in our laboratory for studying the induction of DNA damage in human lung cancer cells after differing doses of the cytosine analog 5-aza-2'-deoxycytidine (5-aza-CdR). Finally, some insights into the potential of this assay in cancer research and work in related fields are offered.

摘要

彗星试验是一种用于检测单个细胞中DNA链断裂的灵敏且快速的方法，2009年是该方法于1984年首次被描述的25周年。随着时间推移，此方法不断改进，但仍未完全标准化，目前各种变体广泛应用，尤其侧重于研究和遗传毒理学方面的应用。在此，我们回顾彗星试验的原理，并引用过去25年中专注于该试验的关键研究。此外，我们还展示了一个例子，说明在我们实验室中彗星试验是如何用于研究不同剂量的胞嘧啶类似物5-氮杂-2'-脱氧胞苷（5-aza-CdR）处理后人肺癌细胞中DNA损伤的诱导情况。最后，我们对该试验在癌症研究及相关领域工作中的潜力提出了一些见解。