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6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 4 is essential for p53-null cancer cells

Oncogene volume 36, pages 3287–3299 (2017)Cite this article

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Abstract

The bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase-4 (PFKFB4) controls metabolic flux through allosteric regulation of glycolysis. Here we show that p53 regulates the expression of PFKFB4 and that p53-deficient cancer cells are highly dependent on the function of this enzyme. We found that p53 downregulates PFKFB4 expression by binding to its promoter and mediating transcriptional repression via histone deacetylases. Depletion of PFKFB4 from p53-deficient cancer cells increased levels of the allosteric regulator fructose-2,6-bisphosphate, leading to increased glycolytic activity but decreased routing of metabolites through the oxidative arm of the pentose-phosphate pathway. PFKFB4 was also required to support the synthesis and regeneration of nicotinamide adenine dinucleotide phosphate (NADPH) in p53-deficient cancer cells. Moreover, depletion of PFKFB4-attenuated cellular biosynthetic activity and resulted in the accumulation of reactive oxygen species and cell death in the absence of p53. Finally, silencing of PFKFB4-induced apoptosis in p53-deficient cancer cells in vivo and interfered with tumour growth. These results demonstrate that PFKFB4 is essential to support anabolic metabolism in p53-deficient cancer cells and suggest that inhibition of PFKFB4 could be an effective strategy for cancer treatment.

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Acknowledgements

We thank C. Esnault (LRI) for advice on ChIP and H. Miess for help with Seahorse Analysis, C. Watkins and J. Bee (LRI, BRU) for help with animal experimentation, F. Lassailly, P. Johnson and T. Snoeks (In vivo imaging facility, LRI) for assistance with in vivo imaging and the LRI research services for technical support. We also thank C. Ade and Barbara Bauer (Theodor-Boveri-Institute, Würzburg) for help with RNAseq analysis and histology, Beatrice Dankworth for help with cell line generation and W. Schmitz for insightful discussions. Cancer Research UK, the German Cancer Aid (grant 111917), the German Research Foundation (FOR 2314), and the CRUK-EPSRC Imaging Centre in Cambridge and Manchester (grant 16465) supported this work.

Author contributions

SR and AS conceived the project and wrote the manuscript. SR, JF, IK, CD, AH, SD, BG and SB performed experiments and analysis of results. RM and SW performed the bioinformatic analysis. AB, KMB, NZ and MHR contributed to the study design and data analysis. All authors commented on the manuscript.

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Authors and Affiliations

  1. Gene Expression Analysis Laboratory, Cancer Research UK London Research Institute, London, UK

    S Ros, B Griffiths & A Schulze

  2. Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK

    S Ros & K M Brindle

  3. Theodor-Boveri-Institute, Biocenter, Am Hubland, Würzburg, Germany

    J Flöter, I Kaymak & A Schulze

  4. Adult Stem Cell Laboratory, The Francis Crick Institute, London, UK

    C Da Costa, S Blake & A Behrens

  5. Protein Phosphorylation Unit, de Duve Institute, Université Catholique de Louvain, Brussels, Belgium

    A Houddane & M H Rider

  6. Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland

    S Dubuis & N Zamboni

  7. Bioinformatics and Biostatistics, The Francis Crick Institute, London, UK

    R Mitter

  8. Comprehensive Cancer Center, Core Unit Bioinformatics, Biocenter, Würzburg, Germany

    S Walz

  9. Faculty of Life Sciences and Medicine, King's College London, Guy's Campus, London, UK

    A Behrens

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Correspondence to A Schulze.

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Ros, S., Flöter, J., Kaymak, I. et al. 6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 4 is essential for p53-null cancer cells. Oncogene 36, 3287–3299 (2017). https://doi.org/10.1038/onc.2016.477

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