Drug-induced cell cycle modulation leading to cell-cycle arrest, nuclear mis-segregation, or endoreplication

BMC Cell Biology

Methodology Article

Kenji Ohtawa¹  Atsushi Miyawaki²  Asako Sakaue-Sawano²  Tamiyo Kobayashi³ 

[1] Brain Science Research Division, Brain Science and Life Technology, Research Foundation, 1-28-12 Narimasu, 175-0094, Itabashi, Tokyo, Japan;Life Function and Dynamics, ERATO, JST, 2-1 Hirosawa, 351-0198, Wako-city, Saitama, Japan;Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, 351-0198, Wako-city, Saitama, Japan;MIS Division, Olympus Corp., 2-3 Kuboyama-cho, 192-8512, Hachioji, Tokyo, Japan;

关键词: Etoposide;    Cell Cycle Progression;    Cdk4 Inhibitor;    Tetraploid Cell;    Trophoblast Giant Cell;   

DOI  :  10.1186/1471-2121-12-2

 received in 2010-07-05, accepted in 2011-01-13,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundCancer cell responses to chemotherapeutic agents vary, and this may reflect different defects in DNA repair, cell-cycle checkpoints, and apoptosis control. Cytometry analysis only quantifies dye-incorporation to examine DNA content and does not reflect the biological complexity of the cell cycle in drug discovery screens.ResultsUsing population and time-lapse imaging analyses of cultured immortalized cells expressing a new version of the fluorescent cell-cycle indicator, Fucci (F luorescent U biquitination-based C ell C ycle I ndicator), we found great diversity in the cell-cycle alterations induced by two anticancer drugs. When treated with etoposide, an inhibitor of DNA topoisomerase II, HeLa and NMuMG cells halted at the G2/M checkpoint. HeLa cells remained there, but NMuMG cells then overrode the checkpoint and underwent nuclear mis-segregation or avoided the checkpoint and entered the endoreplication cycle in a drug concentration dependent manner. In contrast, an inhibitor of Cdk4 led to G1 arrest or endoreplication in NMuMG cells depending upon the initial cell-cycle phase of drug exposure.ConclusionsDrug-induced cell cycle modulation varied not only between different cell types or following treatment with different drugs, but also between cells treated with different concentrations of the same drug or following drug addition during different phases of the cell cycle. By combining cytometry analysis with the Fucci probe, we have developed a novel assay that fully integrates the complexity of cell cycle regulation into drug discovery screens. This assay system will represent a powerful drug-discovery tool for the development of the next generation of anti-cancer therapies.

【 授权许可】

Unknown   
© Sakaue-Sawano et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.