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Automated Analysis of Cell Cycle Phase-Specific DNA Damage Reveals Phase-Specific Differences in Cell Sensitivity to Etoposide.

Authors
  • Luzhin, Artem V1
  • Velichko, Artem K1
  • Razin, Sergey V1, 2, 3
  • Kantidze, Omar L1, 3
  • 1 Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334, Russia.
  • 2 Department of Molecular Biology, Lomonosov Moscow State University, Moscow, 119991, Russia.
  • 3 LIA 1066 French-Russian Joint Cancer Research Laboratory, Villejuif, 94805, France. , (France)
Type
Published Article
Journal
Journal of Cellular Biochemistry
Publisher
Wiley (John Wiley & Sons)
Publication Date
Oct 01, 2016
Volume
117
Issue
10
Pages
2209–2214
Identifiers
DOI: 10.1002/jcb.25615
PMID: 27240930
Source
Medline
Keywords
License
Unknown

Abstract

The comet assay is one of the most widely used approaches for detecting DNA damage; generally, it provides information on the cell population-averaged level of DNA damage. Here, we present an automatic technique for easy measurement of standard comet characteristics and an annotation of the cell cycle phase of each comet. The approach includes the modified neutral comet assay and a pipeline for CellProfiler software designed to analyze DNA damage-related characteristics and annotate the cell cycle phase of each comet. Using this technique we have performed cell cycle phase-specific analysis of DNA damage induced by the topoisomerase II poison etoposide and have shown that the sensitivity of cells to this drug dramatically differed according to their cell cycle phase. It became evident from our results that the proposed protocol provides important additional information that often remains hidden in a standard comet analysis of an asynchronous cell population. J. Cell. Biochem. 117: 2209-2214, 2016. © 2016 Wiley Periodicals, Inc.

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