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  • Original Article
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Absence of the Birt–Hogg–Dubé gene product is associated with increased hypoxia-inducible factor transcriptional activity and a loss of metabolic flexibility

Oncogene volume 30, pages 1159–1173 (2011)Cite this article

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Abstract

Under conditions of reduced tissue oxygenation, hypoxia-inducible factor (HIF) controls many processes, including angiogenesis and cellular metabolism, and also influences cell proliferation and survival decisions. HIF is centrally involved in tumour growth in inherited diseases that give rise to renal cell carcinoma (RCC), such as Von Hippel–Lindau syndrome and tuberous sclerosis complex. In this study, we examined whether HIF is involved in tumour formation of RCC in Birt–Hogg–Dubé syndrome. For this, we analysed a Birt–Hogg–Dubé patient-derived renal tumour cell line (UOK257) that is devoid of the Birt–Hogg–Dubé protein (BHD) and observed high levels of HIF activity. Knockdown of BHD expression also caused a threefold activation of HIF, which was not as a consequence of more HIF1α or HIF2α protein. Transcription of HIF target genes VEGF, BNIP3 and CCND1 was also increased. We found nuclear localization of HIF1α and increased expression of VEGF, BNIP3 and GLUT1 in a chromophobe carcinoma from a Birt–Hogg–Dubé patient. Our data also reveal that UOK257 cells have high lactate dehydrogenase, pyruvate kinase and 3-hydroxyacyl-CoA dehydrogenase activity. We observed increased expression of pyruvate dehydrogenase kinase 1 (a HIF gene target), which in turn leads to increased phosphorylation and inhibition of pyruvate dehydrogenase. Together with increased protein levels of GLUT1, our data reveal that UOK257 cells favour glycolytic rather than lipid metabolism (a cancer phenomenon termed the ‘Warburg effect’). UOK257 cells also possessed a higher expression level of the L-lactate influx monocarboxylate transporter 1 and consequently utilized L-lactate as a metabolic fuel. As a result of their higher dependency on glycolysis, we were able to selectively inhibit the growth of these UOK257 cells by treatment with 2-deoxyglucose. This work suggests that targeting glycolytic metabolism may be used therapeutically to treat Birt–Hogg–Dubé-associated renal lesions.

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Abbreviations

AMPK:

Adenosine monophosphate-dependent protein kinase

BHD:

Birt–Hogg–Dubé protein

DTT:

dithiothreitol

G6PD1:

glucose-6-phosphate dehydrogenase 1

HIF:

hypoxia-inducible factor

HLRCC:

hereditary leiomyomatosis-renal cell carcinoma

LDH:

lactate dehydrogenase

mTOR:

mammalian target of rapamycin

MCT:

monocarboxylate transporter

PDH:

pyruvate dehydrogenase

PDK1:

pyruvate dehydrogenase kinase 1

RCC:

renal cell carcinoma

rpS6:

ribosomal protein S6

TSC:

tuberous sclerosis complex

VHL:

Von Hippel-Lindau

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Acknowledgements

This work was supported by The International Cancer Research Career Development Fellowship (No 06-914/915) (to A Tee). The Myrovlytis Trust supports the labs of A Tee, M v Steensel and A Pause and Partnership for Cures supports A Tee and Mv Steensel. Funding was also provided by the Wellcome Trust (No 088032/Z/08/Z) (to S Land) and the Dutch Cancer Society (grants UM2009-4352 to M v Steensel and UM2010-4609 to T Brinkhuizen). M Kamps, A van Marion and V Winnepenninckx (all of the MUMC) provided valuable assistance with immunohistochemistry. Funding was also provided by the Engineering and Physical Sciences Research Council (EP/E008925/1) and the Biotechnology and Biological Sciences Research Council (BB/F002084/1) (to K Baar). We also thank Wales Gene Park.

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Authors and Affiliations

  1. Institute of Medical Genetics, Cardiff University, Heath Park, Cardiff, Wales, UK

    R S Preston, E A Dunlop, K T Harper, D M Davies & A R Tee

  2. University of California, Davis, CA, USA

    A Philp & K Baar

  3. Department of Dermatology and GROW School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands

    T Claessens, L Gijezen, T Brinkhuizen & M A M van Steensel

  4. Goodman Cancer Centre, McGill University, 3655 Sir William Osler Promenade, Montréal, Québec, Canada

    A B Dydensborg & A Pause

  5. Department of Clinical Genetics, VU Medical Center Amsterdam, Amsterdam, The Netherlands

    F H Menko

  6. Division of Medical Sciences, Centre for Cardiovascular and Lung Biology, Ninewells Hospital and Medical School, University of Dundee, Dundee, UK

    S C Land

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  1. R S Preston
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Correspondence to A R Tee.

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Preston, R., Philp, A., Claessens, T. et al. Absence of the Birt–Hogg–Dubé gene product is associated with increased hypoxia-inducible factor transcriptional activity and a loss of metabolic flexibility. Oncogene 30, 1159–1173 (2011). https://doi.org/10.1038/onc.2010.497

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