Induction of endomitosis-like event in HeLa cells following CHK1 inhibitor treatment
| Autor: | Hisao Homma, Hitomi Nojima, Atsushi Kaida, Masahiko Miura |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Mad2 Cell Survival DNA damage Biophysics Mitosis Time-Lapse Imaging Biochemistry Fluorescence Polyploidy HeLa 03 medical and health sciences 0302 clinical medicine Humans Phosphorylation Protein Kinase Inhibitors Molecular Biology Benzodiazepinones biology Chemistry Cell Cycle Cell Biology Cell cycle biology.organism_classification Cell biology Spindle checkpoint 030104 developmental biology 030220 oncology & carcinogenesis Checkpoint Kinase 1 Mad2 Proteins Cell Cycle Kinetics M Phase Cell Cycle Checkpoints Pyrazoles Cytokinesis DNA Damage HeLa Cells |
| Zdroj: | Biochemical and Biophysical Research Communications. 520:492-497 |
| ISSN: | 0006-291X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect