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PCK2 activation mediates an adaptive response to glucose depletion in lung cancer

Oncogene volume 34, pages 1044–1050 (2015)Cite this article

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Abstract

Cancer cells are reprogrammed to utilize glycolysis at high rates, which provides metabolic precursors for cell growth. Consequently, glucose levels may decrease substantially in underperfused tumor areas. Gluconeogenesis results in the generation of glucose from smaller carbon substrates such as lactate and amino acids. The key gluconeogenic enzyme, phosphoenolpyruvate carboxykinase (PEPCK), has been shown to provide metabolites for cell growth. Still, the role of gluconeogenesis in cancer is unknown. Here we show that the mitochondrial isoform of PEPCK (PCK2) is expressed and active in three lung cancer cell lines and in non-small cell lung cancer samples. PCK2 expression and activity were enhanced under low-glucose conditions. PEPCK activity was elevated threefold in lung cancer samples over normal lungs. To track the conversion of metabolites along the gluconeogenesis pathway, lung cancer cell lines were incubated with 13C3-lactate and label enrichment in the phosphoenolpyruvate (PEP) pool was measured. Under low glucose, all three carbons from 13C3-lactate appeared in the PEP pool, further supporting a conversion of lactate to pyruvate, via pyruvate carboxylase to oxaloacetate, and via PCK2 to phosphoenolpyruvate. PCK2 small interfering RNA and the pharmacological PEPCK inhibitor 3-mercaptopicolinate significantly enhanced glucose depletion-induced apoptosis in A549 and H23 cells, but not in H1299 cells. The growth of H23 multicellular spheroids was significantly reduced by 3-mercaptopicolinate. The results of this study suggest that lung cancer cells may utilize at least some steps of gluconeogenesis to overcome the detrimental metabolic situation during glucose deprivation and that in human lung cancers this pathway is activated in vivo.

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Acknowledgements

We are grateful to Prof. Rudolf Zechner, Karl Franzens University, Graz, Austria, for his advice and valuable discussions. The excellent technical help by Astrid Knopf, Elisabeth Pöllitzer and Alexandra Bertsch, and the contribution by Anna Katschnig are highly appreciated. We are grateful to Dr Günther Fauler and Christina Haas, Medical University of Graz, for their support. The study was supported by the Oesterreichische Nationalbank (Anniversary Fund, project number 12713 to HO) and by a research grant from the state government (KL). ALH is funded by Cancer Research UK and Inspire to Live.

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Authors and Affiliations

  1. Division of Pulmonology, Department of Internal Medicine, Medical University of Graz, Graz, Austria

    K Leithner, A Hrzenjak, C Wohlkoenig & H Olschewski

  2. Ludwig Boltzmann Institute for Lung Vascular Research, Graz, Austria

    A Hrzenjak & A Olschewski

  3. Core Facility Mass Spectrometry and Lipidomics, Center for Medical Research, Medical University of Graz, Graz, Austria

    M Trötzmüller & H C Köfeler

  4. Institute of Molecular Biosciences, Karl Franzens University, Graz, Austria

    T Moustafa

  5. Institute of Pathology, Medical University of Graz, Graz, Austria

    E Stacher

  6. Division of Thoracic and Hyperbaric Surgery, Department of Surgery, Medical University of Graz, Graz, Austria

    J Lindenmann

  7. Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK

    A L Harris

  8. Experimental Anesthesiology, University Clinic for Anesthesiology and Intensive Care Medicine, Medical University of Graz, Graz, Austria

    A Olschewski

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Leithner, K., Hrzenjak, A., Trötzmüller, M. et al. PCK2 activation mediates an adaptive response to glucose depletion in lung cancer. Oncogene 34, 1044–1050 (2015). https://doi.org/10.1038/onc.2014.47

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