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Analysis of cyclin B1 and CDK activity during apoptosis induced by camptothecin treatment

Oncogene volume 25, pages 7361–7372 (2006)Cite this article

Abstract

We have studied the role of cyclins and cyclin-dependent kinase (CDK) activity in apoptosis induced by camptothecin (CPT). In this model, 22% of the cells stain for annexin-V at 24 h and then proceed to be 93% positive by 72 h. This time window permits the analysis of cyclins in cells that are committed to apoptosis but not yet dead. We provide evidence that cyclin protein levels and then associated kinase levels increase after CPT treatment. Strikingly, cyclin B1 and cyclin E1 proteins are present at the same time in CPT treated HT29 cells. Although cyclin B1 and E1 CDK complexes are activated in CPT treated cells, only the cyclin B1 complex is required for apoptosis since reduction of cyclin B1 by RNAi or roscovitine treatment reduces the number of annexin-V-stained cells. We have detected poorly organized chromosomes and phosphorylated histone H3 epitopes at the time of maximum cyclin B1/CDK kinase activity in CPT-treated cells, which suggests that these cells enter a mitotic catastrophe. Understanding which CDKs are required for apoptosis may allow us to better adapt CDK inhibitors for use as anti-cancer compounds.

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Abbreviations

CDK:

cyclin-dependent kinase

CPT:

camptothecin

FCS:

fetal calf serum

PI:

propidium iodide

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Acknowledgements

This work was supported by the Institut de Recherches Servier as part of a program ‘Alliance Stratégique’ with the CNRS. We thank our colleagues in the Cancer Drug Discovery division at the IdRS for valuable discussions and Marie Knockaert for pg beads and advice.

Author information

Author notes

  1. A Borgne

    Present address: Theraptosis, Pasteur Biotop, Institut Pasteur, 25-28, rue du Docteur Roux, 75015, Paris, France

  2. I Versteege

    Present address: Crucell Holland B.V., Archimedesweg 4, PO Box 2048, Leiden, The Netherlands

  3. A Borgne and I Versteege: These authors contributed equally to this study.

Authors and Affiliations

  1. Institut de Recherches Servier, Cancer Drug Discovery, Croissy-sur-Seine, France

    A Borgne, I Versteege, M Mahé, A Studeny, S Léonce, J A Hickman & R M Golsteyn

  2. Institut de Recherches Servier, Molecular Pharmacology and Physiopathology, Croissy-sur-Seine, France

    I Naime & M Rodriguez

  3. CNRS, Station Biologique, Roscoff, France

    L Meijer

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Correspondence to R M Golsteyn.

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Borgne, A., Versteege, I., Mahé, M. et al. Analysis of cyclin B1 and CDK activity during apoptosis induced by camptothecin treatment. Oncogene 25, 7361–7372 (2006). https://doi.org/10.1038/sj.onc.1209718

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