RRC ID 58096
著者 Homma H, Nojima H, Kaida A, Miura M.
タイトル Induction of endomitosis-like event in HeLa cells following CHK1 inhibitor treatment.
ジャーナル Biochem Biophys Res Commun
Abstract The effects of CHK1 inhibitor on cell cycle kinetics have not been fully investigated yet. In this study, we closely analyzed this kinetics using a CHK1 inhibitor (PF00477736) in HeLa cells expressing fluorescent ubiquitination-based cell cycle indicator (Fucci). This system allowed us to visualize cell cycle progression following CHK1 inhibitor treatment in real-time. FACS analysis showed that high levels of DNA damage as determined by γH2AX immunostaining was induced in S phase and that polyploid cells harboring the same levels of DNA damage appeared thereafter. Surprisingly, time-lapse imaging of Fucci fluorescence revealed that many cells entered M phase at once and exhibited prolonged mitosis; eventually progressing to G1 phase not accompanied by cytokinesis; this is an endomitosis-like event. Most of these cells then underwent S/G2 phases at least once, which corroborated the appearance of polyploid cells. However, a small fraction of cells with 2 N DNA content still remained 24 h after the treatment. When co-treated with MAD2 inhibitor, a core factor constituting spindle checkpoint, the 2 N DNA cell fraction disappeared and almost all cells exhibited endomitosis, leading to enhanced sensitivity. Detailed cell cycle analysis revealed that induction of an endomitosis-like event might be associated with CHK1 inhibitor-induced cell death in HeLa cells.
巻・号 520(2)
ページ 492-497
公開日 2019-12-3
DOI 10.1016/j.bbrc.2019.09.046
PII S0006-291X(19)31767-X
PMID 31610912
MeSH Benzodiazepinones / pharmacology* Cell Cycle / drug effects Cell Survival Checkpoint Kinase 1 / antagonists & inhibitors* Checkpoint Kinase 1 / metabolism* DNA Damage Fluorescence HeLa Cells Humans M Phase Cell Cycle Checkpoints / drug effects Mad2 Proteins / antagonists & inhibitors Mad2 Proteins / metabolism Mitosis / drug effects* Phosphorylation / drug effects Polyploidy Protein Kinase Inhibitors / pharmacology* Pyrazoles / pharmacology* Time-Lapse Imaging
IF 2.985
引用数 1
リソース情報
ヒト・動物細胞 HeLa/Fucci(RCB2812)