DNA double-strand breaks and Aurora B mislocalization induced by exposure of early mitotic cells to H2O2appear to increase chromatin bridges and resultant cytokinesis failure.
Aneuploidy, an abnormal number of chromosomes that is a hallmark of cancer cells, can arise from tetraploid/binucleated cells through a failure of cytokinesis. Reactive oxygen species (ROS) have been implicated in various diseases, including cancer. However, the nature and role of ROS in cytokinesis progression and related mechanisms has not been clearly elucidated. Here, using time-lapse analysis of asynchronously growing cells and immunocytochemical analyses of synchronized cells, we found that hydrogen peroxide (H2O2) treatment at early mitosis (primarily prometaphase) significantly induced cytokinesis failure. Cytokinesis failure and the resultant formation of binucleated cells containing nucleoplasmic bridges (NPBs) seemed to be caused by increases in DNA double-strand breaks (DSBs) and subsequent unresolved chromatin bridges. We further found that H2O2induced mislocalization of Aurora B during mitosis. All of these effects were attenuated by pretreatment with N-acetyl-L-cysteine (NAC) or overexpression of Catalase. Surprisingly, the PARP inhibitor PJ34 also reduced H2O2-induced Aurora B mislocalization and binucleated cell formation. Results of parallel experiments with etoposide, a topoisomerase IIα inhibitor that triggers DNA DSBs, suggested that both DNA DSBs and Aurora B mislocalization contribute to chromatin bridge formation. Aurora B mislocalization also appeared to weaken the "abscission checkpoint". Finally, we showed that KRAS-induced binucleated cell formation appeared to be also H2O2-dependent. In conclusion, we propose that a ROS, mainly H2O2increases binucleation through unresolved chromatin bridges caused by DNA damage and mislocalization of Aurora B, the latter of which appears to augment the effect of DNA damage on chromatin bridge formation.
Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon 443-721, South Korea; Genomic Instability Research Center, Ajou University School of Medicine, Suwon 443-721, South Korea; Department of Biomedical Science, Graduate School of Ajou university, Suwon 443-721, South Korea. Electronic address: broselack@naver.com.
Min-Guk
MG
Cho
Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon 443-721, South Korea; Genomic Instability Research Center, Ajou University School of Medicine, Suwon 443-721, South Korea; Department of Biomedical Science, Graduate School of Ajou university, Suwon 443-721, South Korea. Electronic address: hyune20@gmail.com.
Ju-Hyun
JH
Ahn
Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon 443-721, South Korea; Genomic Instability Research Center, Ajou University School of Medicine, Suwon 443-721, South Korea; Department of Biomedical Science, Graduate School of Ajou university, Suwon 443-721, South Korea. Electronic address: heesongchoi@hanmail.net.
Hee-Song
HS
Choi
Department of Biochemistry and Molecular Biology, Ajou University School of Medicine, Suwon 443-721, South Korea; Genomic Instability Research Center, Ajou University School of Medicine, Suwon 443-721, South Korea; Department of Biomedical Science, Graduate School of Ajou university, Suwon 443-721, South Korea. Electronic address: jhlee64@ajou.ac.kr.
Jae-Ho
JH
Lee
D000111:Acetylcysteine / Q000494:pharmacology; D064107:Aurora Kinase B / Q000378:metabolism*; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D048749:Cytokinesis; D053903:DNA Breaks, Double-Stranded; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008938:Mitosis; D010616:Phenanthrenes / Q000494:pharmacology; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
C117307:KRAS protein, human; C434926:N-(oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide hydrochloride; D010616:Phenanthrenes; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D002374:Catalase; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; C576921:AURKB protein, human; D064107:Aurora Kinase B; D016283:Proto-Oncogene Proteins p21(ras); D000111:Acetylcysteine
Aurora B; Binucleation; Chromatin bridges; Cytokinesis failure; H(2)O(2); ROS