Ferroportin and Iron Regulation in Breast Cancer Progression and Prognosis
Science Translational Medicine
4 Aug 2010
Vol 2, Issue 43
p. 43ra56
Iron: Out of Control in Cancer
Toddlers cause an awful lot of damage if let loose without an adult to control their behavior. The same is true for the body’s iron, which when released from its usual protein-binding partners can wreak toxic havoc. Thus, cells usually have an elaborate regulatory system to keep this essential but dangerous metal under control—except for some cancer cells, which have too little of an iron export protein called ferroportin. Pinnix et al. now show that breast cancers with low concentrations of ferroportin tend to contain extra intracellular free iron and to show signs of being more aggressive, suggesting that abnormal iron regulation in cancers may contribute to malignancy.
In an array of normal and cancerous breast epithelial cells, the authors showed that cells with less ferroportin and more of a ferroportin regulator called hepcidin tended to be aggressive. To confirm that the lower ferroportin and higher intracellular free iron actually affected tumor growth, they implanted mice with cancer cells identical except for the addition of extra ferroportin to bring the level of ferroportin in cancer cells to a level near that of normal breast cells. The additional ferroportin in this set of tumors inhibited their growth. In patients, too, the ferroportin status of breast cancer cells proved informative. Women who had breast cancers with lower ferroportin survived for shorter times, and the ferroportin concentrations of a particular tumor could be used to predict survival, providing information beyond the existing markers used clinically such as the estrogen receptor. The authors also came to a more positive conclusion: Having breast cancer with high ferroportin concentrations is a promising sign for a patient and predicts a 90% 10-year survival rate.
Ferroportin, as a marker for iron regulation, may be useful prognostically and in treatment planning. Although it is not yet apparent exactly how higher concentrations of free iron may cause cancer cells to be more aggressive, the cellular iron-regulatory system is clearly a key to understanding breast, and possibly other, cancers. When iron gets out of control, it seems to cause quite a bit of trouble.
Abstract
Ferroportin and hepcidin are critical proteins for the regulation of systemic iron homeostasis. Ferroportin is the only known mechanism for export of intracellular non–heme-associated iron; its stability is regulated by the hormone hepcidin. Although ferroportin profoundly affects concentrations of intracellular iron in tissues important for systemic iron absorption and trafficking, ferroportin concentrations in breast cancer and their influence on growth and prognosis have not been examined. We demonstrate here that both ferroportin and hepcidin are expressed in cultured human breast epithelial cells and that hepcidin regulates ferroportin in these cells. Further, ferroportin protein is substantially reduced in breast cancer cells compared to nonmalignant breast epithelial cells; ferroportin protein abundance correlates with metabolically available iron. Ferroportin protein is also present in normal human mammary tissue and markedly decreased in breast cancer tissue, with the highest degree of anaplasia associated with lowest ferroportin expression. Transfection of breast cancer cells with ferroportin significantly reduces their growth after orthotopic implantation in the mouse mammary fat pad. Gene expression profiles in breast cancers from >800 women reveal that decreased ferroportin gene expression is associated with a significant reduction in metastasis-free and disease-specific survival that is independent of other breast cancer risk factors. High ferroportin and low hepcidin gene expression identifies an extremely favorable cohort of breast cancer patients who have a 10-year survival of >90%. Ferroportin is a pivotal protein in breast biology and a strong and independent predictor of prognosis in breast cancer.
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Supplementary Material
Summary
Materials and Methods
Fig. S1. Ferroportin mRNA and splice variants in breast epithelial cells.
Fig. S2. Assessment of hepcidin mRNA in breast cells by RT-PCR.
Fig. S3. Increased ferroportin expression decreases ferritin in breast cancer cells.
Fig. S4. Relative concentrations of ferroportin protein in MDA-MB-231 cells and transfectants and specific detection of ferroportin by Western blotting.
Fig. S5. Ferroportin protein is decreased in breast cancer.
Fig. S6. Signal intensities of ferroportin and hepcidin relative to control genes and other breast cancer genes in the Uppsala cohort.
Table S1. Ferroportin and hepcidin expression predict outcome in multivariate analyses.
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Published In
Science Translational Medicine
Volume 2 | Issue 43
August 2010
August 2010
Copyright
Copyright © 2010, American Association for the Advancement of Science.
Submission history
Received: 30 March 2010
Accepted: 14 July 2010
Acknowledgments
Acknowledgments: We thank I. De Domenico and J. Kaplan for contributing the ferroportin expression clone and J. Buss for assistance in developing the calcein assay. Funding: Supported in part by grant R37DK42412 from the National Institute of Diabetes and Digestive and Kidney Diseases (F.M.T.), R01 DK071892 (S.V.T.), and by a minority supplement to R37DK42412 (Z.K.P.). Author contributions: Z.K.P. and L.D.M. performed the research and helped write the paper; W.W., T.K., M.C.W., H.H., L.T., and G.S. performed the research; R.D. analyzed the results; S.V.T. and F.M.T. provided funding, designed the research, and wrote the paper. Competing interests: The authors declare that they have no competing interests. Wake Forest University Health Sciences is planning to submit a patent application based on this work. Accession numbers: Accession numbers for the gene expression data are provided in the Microarray data set section of Materials and Methods.
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