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Antiapoptotic effects of roscovitine on camptothecin-induced DNA damage in neuroblastoma cells

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Abstract

In the present study dopaminergic neuroblastoma B65 cells were exposed to Camptothecin (CPT) (0.5–10 μM), either alone or in the presence of roscovitine (ROSC). The results show that CPT induces apoptosis through the activation of ataxia telangiectasia mutated (ATM)-induced cell-cycle alteration in neuroblastoma B65 cells. The apoptotic process is mediated through the activation of cystein proteases, namely calpain/caspases. However, whereas a pan-caspase inhibitor, zVADfmk, inhibited CPT-mediated apoptosis, a calpain inhibitor, calpeptin, did not prevent cell death. Interestingly, CPT also induces CDK5 activation and ROSC (25 μM) blocked CDK5, ATM activation and apoptosis (as measured by caspase-3 activation). By contrast, selective inhibition of ATM, by KU55933, and non-selective inhibition, by caffeine, did not prevent CPT-mediated apoptosis. Thus, we conclude that CDK5 is activated in response to DNA damage and that CDK5 inhibition prevents ATM and p53ser15 activation. However, pharmacological inhibition of ATM using KU55933 and caffeine suggests that ATM inhibition by ROSC is not the only mechanism that might explain the anti-apoptotic effects of this drug in this apoptosis model. Our findings have a potential clinical implication, suggesting that combinatory drugs in the treatment of cancer activation should be administered with caution.

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Acknowledgments

This study was supported by grants from Spain’s “Ministerio de Educación y Ciencia” SAF2009-13093 and BFU2010-19119/BFI the “Fondo de Investigación Sanitaria”, and the “Instituto de Salud Carlos III” PI080400 and PS09/01789 (FEDER FOUNDS). 610RT0405 from Programa Iberoamericano de Ciencia y Tecnologia para el Desarrollo (CYTED). We would like to thank the “Generalitat de Catalunya” for supporting the research groups (2009/SGR00853) and the “Fundació la Marató TV3” (063230). We wish to thank the Language Assessment Service of the University of Barcelona for revising the manuscript.

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Authors and Affiliations

  1. Centros de Investigación Biomédica en Red Enfermedades Neurodegenerativas (CIBERNED), Unitat de Farmacologia i Farmacognòsia. Facultat de Farmàcia, Institut de Biomedicina (IBUB), Universitat de Barcelona, Nucli Universitari de Pedralbes, 08028, Barcelona, Spain

    Javier G. Pizarro, Felix Junyent, Mercè Pallàs & Antoni Camins

  2. Unitat de Bioquímica, Facultat de Medicina i Ciències de la Salut, Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED), Universitat Rovira i Virgili, C/St. Llorenç 21, 43201, Reus (Tarragona), Spain

    Jaume Folch & Felix Junyent

  3. Departamento de Biología Celular y Molecular, CUCBA, Universidad de Guadalajara and División de Neurociencias, Centro de Investigación Biomédica de Occidente, (CIBO), Instituto Mexicano del Seguro Social (IMSS), Sierra Mojada 800, Col. Independencia, , 44340, Guadalajara, Jalisco, Mexico

    Carlos Beas-Zarate

  4. Departament de Biologia Cellular, Facultat de Biologia, Universitat de Barcelona, Barcelona, Spain

    Ester Verdaguer & Carme Auladell

Authors
  1. Javier G. Pizarro
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  2. Jaume Folch
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  8. Antoni Camins
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Correspondence to Antoni Camins.

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Pizarro, J.G., Folch, J., Junyent, F. et al. Antiapoptotic effects of roscovitine on camptothecin-induced DNA damage in neuroblastoma cells. Apoptosis 16, 536–550 (2011). https://doi.org/10.1007/s10495-011-0583-3

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