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Contribution of three-dimensional architecture and tumor-associated fibroblasts to hepcidin regulation in breast cancer

Oncogene (2018) | Download Citation


Hepcidin is a peptide hormone that negatively regulates iron efflux and plays an important role in controlling the growth of breast tumors. In patients with breast cancer, the combined expression of hepcidin and its membrane target, ferroportin, predict disease outcome. However, mechanisms that control hepcidin expression in breast cancer cells remain largely unknown. Here, we use three-dimensional breast cancer spheroids derived from cell lines and breast cancer patients to probe mechanisms of hepcidin regulation in breast cancer. We observe that the extent of hepcidin induction and pathways of its regulation are markedly changed in breast cancer cells grown in three dimensions. In monolayer culture, BMPs, particularly BMP6, regulate hepcidin transcription. When breast cancer cells are grown as spheroids, there is a >10-fold induction in hepcidin transcripts. Microarray analysis combined with knockdown experiments reveal that GDF-15 is the primary mediator of this change. The increase in hepcidin as breast cells develop a three-dimensional architecture increases intracellular iron, as indicated by an increase in the iron storage protein ferritin. Immunohistochemical staining of human breast tumors confirms that both GDF-15 and hepcidin are expressed in breast cancer specimens. Further, levels of GDF-15 are significantly correlated with levels of hepcidin at both the mRNA and protein level in patient samples, consistent with a role for GDF-15 in control of hepcidin in human breast tumors. Inclusion of tumor-associated fibroblasts in breast cancer spheroids further induces hepcidin. This induction is mediated by fibroblast-dependent secretion of IL-6. Breast cancer cells grown as spheroids are uniquely receptive to IL-6-dependent induction of hepcidin by tumor-associated fibroblasts, since IL-6 does not induce hepcidin in cells grown as monolayers. Collectively, our results suggest a new paradigm for tumor-mediated control of iron through the control of hepcidin by tumor architecture and the breast tumor microenvironment.

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This work was supported in part by NCI R01CA188025 (SVT), NCI R01CA171101 (FMT), and NCI F32CA214030 (AK). We thank Li Chen and Drs. Nathaniel Dyment and David Rowe for assistance in image acquisition, Tara L Arvedson (Amgen, Thousand Oaks, CA) for a generous gift of anti-ferroportin antibody and Xiaohong Wang for her assistance in histological sectioning.

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Author notes

    • Daniel Kita

    Present address: Alexion Pharmaceuticals, New Haven, CT, USA

    • David Lemler

    Present address: Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, NC, USA


  1. Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT, USA

    • Nicole Blanchette-Farra
    • , Lia Tesfay
    • , David Lemler
    •  & Suzy V. Torti
  2. Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, USA

    • Daniel Kita
    •  & Kevin P. Claffey
  3. Center for Quantitative Medicine, University of Connecticut Health Center, Farmington, CT, USA

    • Anna Konstorum
  4. Department of Pathology, University of Connecticut Health Center, Farmington, CT, USA

    • Poornima Hegde
  5. Department of Medicine, University of Connecticut Health Center, Farmington, CT, USA

    • Frank M. Torti


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The authors declare that they have no conflict of interest.

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Correspondence to Suzy V. Torti.

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