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Mechanism of leptin expression in breast cancer cells: role of hypoxia-inducible factor-1α

Oncogene volume 27, pages 540–547 (2008)Cite this article

Abstract

We reported previously that the obesity hormone leptin is overexpressed in breast cancer biopsies. Here, we investigated molecular mechanisms involved in this process, focusing on conditions that are associated with obesity, that is, hyperinsulinemia and induction of hypoxia. By using quantitative real-time PCR, immunofluorescent detection of proteins and enzyme-linked immunosorbent assays, we found that treatment of MCF-7 breast cancer cells with high doses of insulin or the hypoxia-mimetic agent CoCl2, or culturing the cells under hypoxic conditions significantly increased the expression of leptin mRNA and protein. Notably, the greatest leptin mRNA and protein expression were observed under combined hyperinsulinemia and hypoxia or hypoxia-mimetic treatments. Luciferase reporter assays suggested that increased leptin synthesis could be related to the activation of the leptin gene promoter. DNA affinity precipitation and chromatin immunoprecipitation experiments revealed that insulin, CoCl2 and/or hypoxia treatments augmented nuclear accumulation of hypoxia-inducible factor-1α (HIF-1α) and increased its interaction with several upstream leptin regulatory sequences, especially with the proximal promoter containing four hypoxia-response elements and three GC-rich regions. By using reverse chromatin precipitation, we determined that loading of HIF-1α on the proximal leptin promoter concurred with the recruitment of p300, the major HIF coactivator, suggesting that the HIF/p300 complex is involved in leptin transcription. The importance of HIF-1α in insulin- and CoCl2-activated leptin mRNA and protein expression was confirmed using RNA interference.

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Acknowledgements

We are grateful to Professor Auwerx, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/Université Louis Pasteur, Illkirch, France, for providing the OB1plasmid. This work was supported by the Sbarro Health Research Organization and the WW Smith Charitable Trust.

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Author notes

  1. S Cascio and V Bartella: These two authors contributed equally to this work.

Authors and Affiliations

  1. Sbarro Institute for Cancer Research and Molecular Medicine, Temple University, Philadelphia, PA, USA

    S Cascio, V Bartella, A Giordano & E Surmacz

  2. Department of Surgical and Oncological Sciences Section of Medical Oncology, University of Palermo, Palermo, Italy

    S Cascio & A Russo

  3. Department of Pharmaco-Biology, University of Calabria, Cosenza, Italy

    V Bartella

  4. Department of Oncology, University of Verona, Italy

    A Auriemma

  5. Department of Pathology and Laboratory Medicine, Drexel University College of Medicine, Philadelphia, PA, USA

    G J Johannes

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  1. S Cascio
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  2. V Bartella
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  4. G J Johannes
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  7. E Surmacz
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Correspondence to E Surmacz.

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Cascio, S., Bartella, V., Auriemma, A. et al. Mechanism of leptin expression in breast cancer cells: role of hypoxia-inducible factor-1α. Oncogene 27, 540–547 (2008). https://doi.org/10.1038/sj.onc.1210660

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