title: Automated Analysis of Cell Cycle Phase-Specific DNA Damage Reveals Phase-Specific Differences in Cell Sensitivity to Etoposide.
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.
authors.affiliation: Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334, Russia.
authors.forename: Artem V
authors.initials: AV
authors.lastname: Luzhin
authors.affiliation: Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334, Russia.
authors.forename: Artem K
authors.initials: AK
authors.lastname: Velichko
authors.affiliation: Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334, Russia.|Department of Molecular Biology, Lomonosov Moscow State University, Moscow, 119991, Russia.|LIA 1066 French-Russian Joint Cancer Research Laboratory, Villejuif, 94805, France.
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Razin
authors.affiliation: Laboratory of Structural and Functional Organization of Chromosomes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, 119334, Russia.|LIA 1066 French-Russian Joint Cancer Research Laboratory, Villejuif, 94805, France.
authors.forename: Omar L
authors.initials: OL
authors.lastname: Kantidze
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D001331:Automation; D015153:Blotting, Western; D002453:Cell Cycle / Q000187:drug effects*; D020552:Comet Assay; D004249:DNA Damage; D005047:Etoposide / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D012984:Software
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D005047:Etoposide
keywords: CELL CYCLE; COMET ASSAY; CellProfiler; DNA DAMAGE; IMAGE ANALYSIS