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SHARP1 suppresses breast cancer metastasis by promoting degradation of hypoxia-inducible factors

Nature volume 487, pages 380–384 (2012)Cite this article

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Abstract

The molecular determinants of malignant cell behaviours in breast cancer remain only partially understood1. Here we show that SHARP1 (also known as BHLHE41 or DEC2) is a crucial regulator of the invasive and metastatic phenotype in triple-negative breast cancer (TNBC), one of the most aggressive types of breast cancer. SHARP1 is regulated by the p63 metastasis suppressor and inhibits TNBC aggressiveness through inhibition of hypoxia-inducible factor 1α (HIF-1α) and HIF-2α (HIFs). SHARP1 opposes HIF-dependent TNBC cell migration in vitro, and invasive or metastatic behaviours in vivo. SHARP1 is required, and sufficient, to limit expression of HIF-target genes. In primary TNBC, endogenous SHARP1 levels are inversely correlated with those of HIF targets. Mechanistically, SHARP1 binds to HIFs and promotes HIF proteasomal degradation by serving as the HIF-presenting factor to the proteasome. This process is independent of pVHL (von Hippel–Lindau tumour suppressor), hypoxia and the ubiquitination machinery. SHARP1 therefore determines the intrinsic instability of HIF proteins to act in parallel to, and cooperate with, oxygen levels. This work sheds light on the mechanisms and pathways by which TNBC acquires invasiveness and metastatic propensity.

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Figure 1: SHARP1 is an inhibitor of HIF activity.
Figure 2: SHARP1 is a suppressor of invasion, migration and metastasis by inhibiting HIFs.
Figure 3: SHARP1 promotes HIF-1α proteasomal degradation independently of oxygen levels and pVHL.
Figure 4: SHARP1 promotes the interaction of HIF-1α with the proteasome.

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Acknowledgements

We thank O. Wessely, G. Del Sal, C. Oakman and A. Di Leo for comments; and W. Kaelin, C. Borner, S. Libutti, Y. Maeda and Y. Kato for gifts of reagents. M.M. is a recipient of a FIRC (Federazione Italiana Ricerca Cancro) fellowship. We are in debt to E. Tagliafico and the Modena Affimetrix platform team for help with microarrays. E.E. is a recipient of a Cariparo PhD fellowship. M.M. was a recipient of an AIRC (Italian Association for Cancer Research) fellowship. This work is supported by a Young Italian Researchers grant of the Italian Welfare Ministry and an AIRC (Associazione Italiana per la Ricerca sul Cancro) MFAG grant to M.C.; a Fondazione Città della Speranza Grant, and MIUR (Ministero dell'Istruzione dell'Università e della Ricerca Italia) and PRIN grants to G.B; and an AIRC Principal Investigator grant, an AIRC Special Program Molecular Clinical Oncology ‘5 per mille’ grant, an HSFP grant, a University of Padua Strategic grant, an IIT Excellence grant, a CNR–Miur Epigenetics Flagship project, and a Comitato Promotore Telethon grant to S.P.

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Authors and Affiliations

  1. Department of Medical Biotechnologies, University of Padua School of Medicine, Viale Colombo 3, 35131 Padua, Italy,

    Marco Montagner, Elena Enzo, Francesca Zanconato, Michelangelo Cordenonsi & Stefano Piccolo

  2. Department of Biomedical Sciences, Center for Genome Research, University of Modena and Reggio Emilia, Via G. Campi 287, 41100 Modena, Italy,

    Mattia Forcato & Silvio Bicciato

  3. Department of Medical Diagnostic Science and Special Therapies, Section of Pathology, University of Padua, Viale Gabelli 2, 35126 Padua, Italy,

    Anna Parenti

  4. Department of Pediatrics, Clinical and Experimental Hematology, University of Padova, Via Giustiniani 3, 35128 Padova, Italy,

    Elena Rampazzo & Giuseppe Basso

  5. Division of Anatomic Pathology, Hospital San Bassiano, Via dei Lotti 40, 36061 Bassano del Grappa, Italy,

    Genesio Leo

  6. Department of Surgery, Oncology and Gastroenterology, and Istituto Oncologico Veneto IRCCS, Via Gattamelata 64, 35126 Padua, Italy,

    Antonio Rosato

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  1. Marco Montagner
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Contributions

M.M., M.C. and S.P. designed research, and M.M., E.E, M.F., M.C., E.R., G.B., G.L. and F.Z. performed experiments. M.C., S.B. and M.F. performed bioinformatics analysis, and A.R. and A.P. helped with assays in mice and tumour pathology. M.M., M.C. and S.P. wrote the manuscript.

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Correspondence to Stefano Piccolo.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information 1

This file contains Supplementary Figures 1-29, Supplementary Methods, Supplementary Tables 1-2, 4, and 6-11 (see separate files for Supplementary Tables 3 and 5) and Supplementary references. (PDF 15593 kb)

Supplementary Table 3

This file contains a table with the Characteristics of the 250 TNBC samples. (XLS 92 kb)

Supplementary Table 5

This file contains a table with the Genes identified by SAM as up-regulated in Sharp1/CyclinG2 Low TNBCs compared to Sharp1/CyclinG2 High TNBCs. (XLS 550 kb)

Supplementary Information 2

This file contains the Sweave document in .pdf format for the analysis of gene expression data from MDA-MB-231 cells depleted of HIF-1α and HIF-2α (MDA-shHIF) and MDA-MB-231 cells overexpressing Sharp1 (MDA-Sharp1). (PDF 272 kb)

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Montagner, M., Enzo, E., Forcato, M. et al. SHARP1 suppresses breast cancer metastasis by promoting degradation of hypoxia-inducible factors. Nature 487, 380–384 (2012). https://doi.org/10.1038/nature11207

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