Abstract

Blocking mitotic progression has been proposed as an attractive therapeutic strategy to impair proliferation of tumour cells. However, how cells survive during prolonged mitotic arrest is not well understood. We show here that survival during mitotic arrest is affected by the special energetic requirements of mitotic cells. Prolonged mitotic arrest results in mitophagy-dependent loss of mitochondria, accompanied by reduced ATP levels and the activation of AMPK. Oxidative respiration is replaced by glycolysis owing to AMPK-dependent phosphorylation of PFKFB3 and increased production of this protein as a consequence of mitotic-specific translational activation of its mRNA. Induction of autophagy or inhibition of AMPK or PFKFB3 results in enhanced cell death in mitosis and improves the anti-tumoral efficiency of microtubule poisons in breast cancer cells. Thus, survival of mitotic-arrested cells is limited by their metabolic requirements, a feature with potential implications in cancer therapies aimed to impair mitosis or metabolism in tumour cells.

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Acknowledgements

We thank M. Soengas (Spanish National Cancer Research Centre, Spain), J. P. Bolaños (Institute of Functional Biology and Genomics, Spain) and N. Mizushima (The University of Tokyo, Japan) for reagents. We also thank S. Velasco for help with the Seahorse apparatus, J. Muñoz and M. P. Ximénez for their support in phospho-proteomics experiments, and F. Escobar and B. Pou for help with electron microscopy. E.D., C.M. and M.S.-R. were supported by the Spanish Fondo de Investigaciones Sanitarias (Madrid), MINECO (Juan de la Cierva programme) and Asociación Española contra el Cáncer (AECC), respectively. L.E.-M. is a recipient of a JAE predoctoral fellowship from the CSIC. A.K.S. was supported by USPHS grants RO1DK19514, RO1DK67509. G.V. was supported by grants from the Spanish Ministry of Economy and Competitiveness (MINECO) and Fondo Europeo de Desarrollo Regional (FEDER) (PI12/02248), Fundació La Marató de TV3 (m12 20134031), and Fundación Mutua Madrileña (AP101042012). M.L. was supported by the European Community’s Seventh Framework Programme under grant agreement no. 281854—the ObERStress (European Research Council project). E.R. was financially supported by a MINECO grant (SAF 2010-20256). Work in the R.M. laboratory was supported by the Fundación Botín, Banco Santander and MINECO (BFU2011-30121, BFU2014-52125-REDT and Consolider RNAREG CSD2009-00080). Work in the P.B. laboratory is supported by a grant from the Spanish Ministry for Economy and Competitiveness (MINECO; SAF2012-36079). Work in the M.M. laboratory was supported by grants from the MINECO (SAF2012-38215), Consolider-Ingenio 2010 Programme (SAF2014-57791-REDC), Excellence Network CellSYS (BFU2014-52125-REDT), the OncoCycle Programme (S2010/BMD-2470) from the Comunidad de Madrid, Worldwide Cancer Research (WCR no. 15-0278), and the European Union Seventh Framework Programme (MitoSys project; HEALTH-F5-2010-241548).

Author information

Affiliations

  1. Cell Division and Cancer Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain

    • Elena Doménech
    • , Carolina Maestre
    • , David Partida
    • , María Salazar-Roa
    •  & Marcos Malumbres
  2. Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, CSIC, E-28040 Madrid, Spain

    • Lorena Esteban-Martínez
    • , Esther Seco
    •  & Patricia Boya
  3. ICREA and Institute for Research in Biomedicine (IRB), Barcelona 08028, Spain

    • Rosa Pascual
    • , Gonzalo Fernández-Miranda
    •  & Raúl Méndez
  4. Spectroscopy and Nuclear Magnetic Resonance Unit, CNIO, Madrid 28029, Spain

    • Ramón Campos-Olivas
  5. Confocal Microscopy Unit, CNIO, Madrid  28029, Spain

    • Manuel Pérez
    •  & Diego Megias
  6. Division of Endocrinology, Diabetes & Nutrition, Boston University School of Medicine, Boston, Massachusetts 02215, USA

    • Katherine Allen
    •  & Asish K. Saha
  7. Department of Physiology, CIMUS, University of Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela 15782, Spain

    • Miguel López
  8. CIBER Fisiopatología de la Obesidad y Nutrición (CIBERobn), 15706, Spain

    • Miguel López
  9. Department of Biochemistry and Molecular Biology I, School of Biology, Complutense University, and Instituto de Investigaciones Sanitarias San Carlos (IdISSC), 28040 Madrid, Spain

    • Guillermo Velasco
  10. Department of Cellular and Molecular Medicine, Centro de Investigaciones Biológicas, CSIC, E-28040 Madrid, Spain

    • Eduardo Rial

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Contributions

E.D. and M.S.-R. performed most of the cellular and biochemical assays with the help of C.M. and D.P. L.E.-M., E.S. and P.B. collaborated in the analysis of mitophagy, and G.F.-M., R.P. and R.M. collaborated in the analysis of translation. R.C.-O. generated the NMR data. M.P. and D.M. helped with microscopy analysis. A.K.S. performed the AMP/ATP measurements and M.L. contributed to the analysis of AMPK. E.R. helped with metabolic measurements. E.D., C.M., G.V., E.R., P.B., M.S.-R. and M.M. analysed the data. M.M. conceived the project, M.S.R. and M.M., supervised the experiments, and M.M. wrote the manuscript with the help of all co-authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to María Salazar-Roa or Marcos Malumbres.

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https://doi.org/10.1038/ncb3231

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