Breast Cancer Research and Treatment

, Volume 125, Issue 2, pp 421–430 | Cite as

Enhanced serine production by bone metastatic breast cancer cells stimulates osteoclastogenesis

  • Sirkku PollariEmail author
  • Sanna-Maria Käkönen
  • Henrik Edgren
  • Maija Wolf
  • Pekka Kohonen
  • Henri Sara
  • Theresa Guise
  • Matthias Nees
  • Olli Kallioniemi
Preclinical study


Since bone metastatic breast cancer is an incurable disease, causing significant morbidity and mortality, an understanding of the underlying molecular mechanisms would be highly valuable. Here, we describe in vitro and in vivo evidences for the importance of serine biosynthesis in the metastasis of breast cancer to bone. We first characterized the bone metastatic propensity of the MDA-MB-231(SA) cell line variant as compared to the parental MDA-MB-231 cells by radiographic and histological observations in the inoculated mice. Genome-wide gene expression profiling of this isogenic cell line pair revealed that all the three genes involved in the l-serine biosynthesis pathway, phosphoglycerate dehydrogenase (PHGDH), phosphoserine aminotransferase 1 (PSAT1), and phosphoserine phosphatase (PSPH) were upregulated in the highly metastatic variant. This pathway is the primary endogenous source for l-serine in mammalian tissues. Consistently, we observed that the proliferation of MDA-MB-231(SA) cells in serine-free conditions was dependent on PSAT1 expression. In addition, we observed that l-serine is essential for the formation of bone resorbing human osteoclasts and may thus contribute to the vicious cycle of osteolytic bone metastasis. High expression of PHGDH and PSAT1 in primary breast cancer was significantly associated with decreased relapse-free and overall survival of patients and malignant phenotypic features of breast cancer. In conclusion, high expression of serine biosynthesis genes in metastatic breast cancer cells and the stimulating effect of l-serine on osteoclastogenesis and cancer cell proliferation indicate a functionally critical role for serine biosynthesis in bone metastatic breast cancer and thereby an opportunity for targeted therapeutic interventions.


Breast cancer Bone metastasis Osteoclast l-serine 



This study was supported by the Academy of Finland Center of Excellence grant (Center of Excellence in Translational Genome-Scale Biology 2006–2011), TIME (Disseminated Tumour Cells as Targets for Inhibiting Metastasis of Epithelial Tumours) project, Drug Discovery Graduate School, NIH grant R01 CA69158 from the National Cancer Institute, and by grants from the Finnish Cancer Organisations, the Sigrid Jusélius Foundation, and the Finnish Cultural Foundation. We thank Barry G. Grubbs and Rami Käkönen for excellent technical assistance in the mouse studies, Pharmatest Services Ltd ( for technical help and discussions regarding the osteoclast cultures, and John Patrick Mpindi for help in bioinformatic analyses.

Supplementary material

10549_2010_848_MOESM1_ESM.pdf (199 kb)
Supplementary material 1 (PDF 199 kb)


  1. 1.
    Vargas SJ, Gillespie MT, Powell GJ et al (1992) Localization of parathyroid hormone-related protein mRNA expression in breast cancer and metastatic lesions by in situ hybridization. J Bone Miner Res 7:971–979PubMedCrossRefGoogle Scholar
  2. 2.
    Guise TA, Yin JJ, Taylor SD et al (1996) Evidence for a causal role of parathyroid hormone-related protein in the pathogenesis of human breast cancer-mediated osteolysis. J Clin Investig 98:1544–1549. doi: 10.1172/JCI118947 PubMedCrossRefGoogle Scholar
  3. 3.
    Akhtari M, Mansuri J, Newman KA et al (2008) Biology of breast cancer bone metastasis. Cancer Biol Ther 7:3–9PubMedCrossRefGoogle Scholar
  4. 4.
    Yin JJ, Selander K, Chirgwin JM et al (1999) TGF-beta signaling blockade inhibits PTHrP secretion by breast cancer cells and bone metastases development. J Clin Investig 103:197–206. doi: 10.1172/JCI3523 PubMedCrossRefGoogle Scholar
  5. 5.
    Kang Y, Siegel PM, Shu W et al (2003) A multigenic program mediating breast cancer metastasis to bone. Cancer Cell 3:537–549PubMedCrossRefGoogle Scholar
  6. 6.
    Yoneda T, Williams PJ, Hiraga T et al (2001) A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro. J Bone Miner Res 16:1486–1495PubMedCrossRefGoogle Scholar
  7. 7.
    Bendre MS, Margulies AG, Walser B et al (2005) Tumor-derived interleukin-8 stimulates osteolysis independent of the receptor activator of nuclear factor-kappaB ligand pathway. Cancer Res 65:11001–11009. doi: 10.1158/0008-5472.CAN-05-2630 PubMedCrossRefGoogle Scholar
  8. 8.
    Kakonen SM, Selander KS, Chirgwin JM et al (2002) Transforming growth factor-beta stimulates parathyroid hormone-related protein and osteolytic metastases via Smad and mitogen-activated protein kinase signaling pathways. J Biol Chem 277:24571–24578. doi: 10.1074/jbc.M202561200 PubMedCrossRefGoogle Scholar
  9. 9.
    R Development Core Team (2004) R: a language and environment for statistical computing. R foundation for statistical computing, Vienna, AustriaGoogle Scholar
  10. 10.
    Gentleman RC, Carey VJ, Bates DM et al (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80. doi: 10.1186/gb-2004-5-10-r80 PubMedCrossRefGoogle Scholar
  11. 11.
    Irizarry RA, Bolstad BM, Collin F et al (2003) Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 31:e15PubMedCrossRefGoogle Scholar
  12. 12.
    Dai M, Wang P, Boyd AD et al (2005) Evolving gene/transcript definitions significantly alter the interpretation of GeneChip data. Nucleic Acids Res 33:e175. doi: 10.1093/nar/gni179 PubMedCrossRefGoogle Scholar
  13. 13.
    Miller LD, Smeds J, George J et al (2005) An expression signature for p53 status in human breast cancer predicts mutation status, transcriptional effects, and patient survival. Proc Natl Acad Sci USA 102:13550–13555. doi: 10.1073/pnas.0506230102 PubMedCrossRefGoogle Scholar
  14. 14.
    Sorlie T, Tibshirani R, Parker J et al (2003) Repeated observation of breast tumor subtypes in independent gene expression data sets. Proc Natl Acad Sci USA 100:8418–8423. doi: 10.1073/pnas.0932692100 PubMedCrossRefGoogle Scholar
  15. 15.
    Dennis G Jr, Sherman BT, Hosack DA et al (2003) DAVID: database for annotation, visualization, and integrated discovery. Genome Biol 4:P3PubMedCrossRefGoogle Scholar
  16. 16.
    Zhang B, Kirov S, Snoddy J (2005) WebGestalt: an integrated system for exploring gene sets in various biological contexts. Nucleic Acids Res 33:W741–W748. doi: 10.1093/nar/gki475 PubMedCrossRefGoogle Scholar
  17. 17.
    Lewis RM, Glazier J, Greenwood SL et al (2007) l-serine uptake by human placental microvillous membrane vesicles. Placenta 28:445–452. doi: 10.1016/j.placenta.2006.06.014 PubMedCrossRefGoogle Scholar
  18. 18.
    Ogawa T, Ishida-Kitagawa N, Tanaka A et al (2006) A novel role of l-serine (l-Ser) for the expression of nuclear factor of activated T cells (NFAT)2 in receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis in vitro. J Bone Miner Metab 24:373–379. doi: 10.1007/s00774-006-0705-0 PubMedCrossRefGoogle Scholar
  19. 19.
    Alatalo SL, Halleen JM, Hentunen TA et al (2000) Rapid screening method for osteoclast differentiation in vitro that measures tartrate-resistant acid phosphatase 5b activity secreted into the culture medium. Clin Chem 46:1751–1754PubMedGoogle Scholar
  20. 20.
    Snell K (1984) Enzymes of serine metabolism in normal, developing and neoplastic rat tissues. Adv Enzyme Regul 22:325–400PubMedCrossRefGoogle Scholar
  21. 21.
    Arriza JL, Kavanaugh MP, Fairman WA et al (1993) Cloning and expression of a human neutral amino acid transporter with structural similarity to the glutamate transporter gene family. J Biol Chem 268:15329–15332PubMedGoogle Scholar
  22. 22.
    Broer A, Wagner C, Lang F et al (2000) Neutral amino acid transporter ASCT2 displays substrate-induced Na+ exchange and a substrate-gated anion conductance. Biochem J 346(Pt 3):705–710PubMedCrossRefGoogle Scholar
  23. 23.
    Chaudhry FA, Schmitz D, Reimer RJ et al (2002) Glutamine uptake by neurons: interaction of protons with system a transporters. J Neurosci 22:62–72PubMedGoogle Scholar
  24. 24.
    Fukasawa Y, Segawa H, Kim JY et al (2000) Identification and characterization of a Na(+)-independent neutral amino acid transporter that associates with the 4F2 heavy chain and exhibits substrate selectivity for small neutral d- and l-amino acids. J Biol Chem 275:9690–9698PubMedCrossRefGoogle Scholar
  25. 25.
    de Koning TJ, Snell K, Duran M et al (2003) l-serine in disease and development. Biochem J 371:653–661. doi: 10.1042/BJ20021785 PubMedCrossRefGoogle Scholar
  26. 26.
    Martens JW, Nimmrich I, Koenig T et al (2005) Association of DNA methylation of phosphoserine aminotransferase with response to endocrine therapy in patients with recurrent breast cancer. Cancer Res 65:4101–4117. doi: 10.1158/0008-5472.CAN-05-0064 PubMedCrossRefGoogle Scholar
  27. 27.
    Kozlow W, Guise TA (2005) Breast cancer metastasis to bone: mechanisms of osteolysis and implications for therapy. J Mammary Gland Biol Neoplasia 10:169–180. doi: 10.1007/s10911-005-5399-8 PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Sirkku Pollari
    • 1
    Email author
  • Sanna-Maria Käkönen
    • 2
    • 5
  • Henrik Edgren
    • 3
  • Maija Wolf
    • 3
  • Pekka Kohonen
    • 1
  • Henri Sara
    • 1
  • Theresa Guise
    • 4
  • Matthias Nees
    • 1
  • Olli Kallioniemi
    • 3
  1. 1.Medical BiotechnologyVTT Technical Research Center of Finland and Turku Center for BiotechnologyTurkuFinland
  2. 2.Institute of Biomedicine, Department of AnatomyUniversity of TurkuTurkuFinland
  3. 3.Institute for Molecular Medicine Finland (FIMM)University of HelsinkiHelsinkiFinland
  4. 4.Division of EndocrinologyIndiana UniversityIndianapolisUSA
  5. 5.Amgen AB FinlandEspooFinland

Personalised recommendations