Clinical & Experimental Metastasis

, Volume 29, Issue 4, pp 327–338 | Cite as

Augmented autocrine bone morphogenic protein (BMP) 7 signaling increases the metastatic potential of mouse breast cancer cells

  • Hirofumi Sakai
  • Mutsuo Furihata
  • Chie Matsuda
  • Munehisa Takahashi
  • Hiroshi Miyazaki
  • Takeo Konakahara
  • Toru Imamura
  • Tomoko Okada
Research Paper


As malignant breast cancers progress, they acquire the ability to spread to other regions of the body, including bone and lung, but the molecular mechanism underlying the increase in metastatic potential is not fully understood. Here we studied murine 4T1E/M3 highly bone marrow metastatic breast cancer cells, which we established previously. These cells show upregulated expression of bone morphogenetic protein (BMP) 7 and BMP receptors, as well as augmented phosphorylation of Smad1/5/8. Both anchorage-independent cell growth measured in colony forming assays and cell migration measured in wound healing assays were suppressed in 4T1E/M3 cells following treatment with a neutralizing anti-BMP7 antibody or knockdown of BMP7 gene expression. In addition, metastasis of 4T1E/M3 cells to the spine and lung and intracellular levels of phosphorylated Smad1/5/8 were suppressed by knocking down BMP7. Conversely, overexpression of BMP7 in the weakly metastatic parental 4T1E cells augmented their anchorage-independent growth, migration and metastasis to spine and lung. Taken together, our results strongly suggest that augmented autocrine BMP7 signaling leads to increases in the anchorage-independent cell growth, migration and metastatic potential in our bone marrow metastatic breast cancer model.


Breast caner Metastasis BMP7 Anchorage-independent growth Migration 



We thank Ms. Yoko Ezaki for her technical assistance. This work was supported in part by the Japan Science and Technology Agency (JST).


  1. 1.
    Takahashi M, Furihata M, Akimitsu N, Watanabe M, Kaul S, Yumoto N, Okada T (2008) A highly bone marrow metastatic murine breast cancer model established through in vivo selection exhibits enhanced anchorage-independent growth and cell migration mediated by ICAM-1. Clin Exp Metastasis 25:517–529PubMedCrossRefGoogle Scholar
  2. 2.
    Takahashi M, Miyazaki H, Furihata M, Sakai H, Konakahara T, Watanabe M, Okada T (2009) Chemokine CCL2/MCP-1 negatively regulates metastasis in a highly bone marrow-metastatic mouse breast cancer model. Clin Exp Metastasis 26:817–828PubMedCrossRefGoogle Scholar
  3. 3.
    Chen D, Zhao M, Mundy GR (2004) Bone morphogenetic proteins. Growth Factors 4:233–241CrossRefGoogle Scholar
  4. 4.
    Ye L, Lewis-Russell JM, Kyanaston HG, Jiang WG (2007) Bone morphogenetic proteins and their receptor signaling in prostate cancer. Histol Histopathol 10:1129–1147Google Scholar
  5. 5.
    Asahina I, Sampath TK, Hauschka PV (1996) Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells. Exp Cell Res 222:38–47PubMedCrossRefGoogle Scholar
  6. 6.
    Autzen P, Robson CN, Bjartell A, Malcolm AJ, Johnson MI, Neal DE, Hamdy FC (1998) Bone morphogenetic protein 6 in skeletal metastases from prostate cancer and other common human malignancies. Br J Cancer 78:1219–1223PubMedCrossRefGoogle Scholar
  7. 7.
    Masuda H, Fukabori Y, Nakano K, Takezawa Y, CSuzuki T, Yamanaka H (2003) Increased expression of bone morphogenetic protein-7 in bone metastatic prostate cancer. Prostate 54:268–274PubMedCrossRefGoogle Scholar
  8. 8.
    Miyazono K, Kamiya Y, Morikawa M (2010) Bone morphogenetic protein receptors and signal transduction. J Biochem 147:35–51PubMedCrossRefGoogle Scholar
  9. 9.
    Miyazawa K, Shinozaki M, Hara T, Furuya T, Miyazono K (2002) Two major Smad pathways in TGF-beta superfamily signalling. Genes Cells 12:1191–1204CrossRefGoogle Scholar
  10. 10.
    Massagué J, Blain SW, Lo RS (2000) TGFbeta signaling in growth control, cancer, and heritable disorders. Cell 103:295–309PubMedCrossRefGoogle Scholar
  11. 11.
    Asada M, Honda E, Imamura T (2006) Construction of pcDNA 3.1-based vectors with blasticidin and puromycin resistant markers. Anal Biochem 352:305–307PubMedCrossRefGoogle Scholar
  12. 12.
    Alarmo EL, Pärssinen J, Ketolainen JM, Savinainen K, Karhu R, Kallioniemi A (2009) BMP7 influences proliferation, migration, and invasion of breast cancer cells. Cancer Lett 275:35–43PubMedCrossRefGoogle Scholar
  13. 13.
    Ichikawa T, Suenaga Y, Koda T, Ozaki T, Nakagawara A (2008) DeltaNp63/BMP-7-dependent expression of matrilin-2 is involved in keratinocyte migration in response to wounding. Biochem Biophys Res Commun 369:994–1000PubMedCrossRefGoogle Scholar
  14. 14.
    Rothhammer T, Braig S, Bosserhoff AK (2008) Bone morphogenetic proteins induce expression of metalloproteinases in melanoma cells and fibroblasts. Eur J Cancer 44:2526–2534PubMedCrossRefGoogle Scholar
  15. 15.
    Zeisberg M, Hanai J, Sugimoto H, Mammoto T, Charytan D, Strutz F, Kalluri R (2003) BMP-7 counteracts TGF-beta1-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat Med 9:964–968PubMedCrossRefGoogle Scholar
  16. 16.
    Rees JR, Onwuegbusi BA, Save VE, Alderson D, Fitzgerald RC (2006) In vivo and in vitro evidence for transforming growth factor-beta1-mediated epithelial to mesenchymal transition in esophageal adenocarcinoma. Cancer Res 66:9583–9590PubMedCrossRefGoogle Scholar
  17. 17.
    Yang S, Zhong C, Frenkel B, Reddi AH, Roy-Burman P (2005) Diverse biological effect and Smad signaling of bone morphogenetic protein 7 in prostate tumor cells. Cancer Res 65:5769–5777PubMedCrossRefGoogle Scholar
  18. 18.
    Rothhammer T, Poser I, Soncin F, Bataille F, Moser M, Bosserhoff AK (2005) Bone morphogenic proteins are overexpressed in malignant melanoma and promote cell invasion and migration. Cancer Res 65:448–456PubMedGoogle Scholar
  19. 19.
    Alarmo EL, Korhonen T, Kuukasjärvi T, Huhtala H, Holli K, Kallioniemi A (2008) Bone morphogenetic protein 7 expression associates with bone metastasis in breast carcinomas. Ann Oncol 19:308–314PubMedCrossRefGoogle Scholar
  20. 20.
    Motoyama K, Tanaka F, Kosaka Y, Mimori K, Uetake H, Inoue H, Sugihara K, Mori M (2008) Clinical significance of BMP7 in human colorectal cancer. Ann Surg Oncol 15:1530–1537PubMedCrossRefGoogle Scholar
  21. 21.
    Rothhammer T, Wild PJ, Meyer S, Bataille F, Pauer A, Klinkhammer-Schalke M, Hein R, Hofstaedter F, Bosserhoff AK (2007) Bone morphogenetic protein 7 (BMP7) expression is a potential novel prognostic marker for recurrence in patients with primary melanoma. Cancer Biomark 3:111–117PubMedGoogle Scholar
  22. 22.
    Buijs JT, Henriquez NV, van Overveld PG (2007) Bone morphogenetic protein 7 in the development and treatment of bone metastases from breast cancer. Cancer Res 67:8742–8751PubMedCrossRefGoogle Scholar
  23. 23.
    Na YR, Seok SH, Kim DJ et al (2009) Bone morphogenetic protein 7 induces mesenchymal -to-epithelial transition in melanoma cells, leading to inhibition of metastasis. Cancer Sci 100:2218–2225PubMedCrossRefGoogle Scholar
  24. 24.
    Chen J, Ye L, Xie F, Yang Y, Zhang L, Jiang WG (2010) Expression of bone morphogenetic protein 7 in lung cancer and its biological impact on lung cancer cells. Anticancer Res 4:1113–1120Google Scholar
  25. 25.
    Uruno T, Oki J, Ozawa K, Miyakawa K, Ueno H, Imamura T (1999) Distinct regulation of myoblast differentiation by intracellular and extracellular fibroblast growth factor-1. Growth Factors 17:93–113PubMedCrossRefGoogle Scholar
  26. 26.
    Schoner W, Scheiner-Bobis G (2007) Endogenous and exogenous cardiac glycosides: their roles in hypertension, salt metabolism, and cell growth. Am J Physiol Cell Physiol 293:C509–C536PubMedCrossRefGoogle Scholar
  27. 27.
    Fournier PG, Guise TA (2007) BMP7: a new bone metastases prevention? Am J Pathol 171:739–743PubMedCrossRefGoogle Scholar
  28. 28.
    Morrissey C, Brown LG, Pitts TE, Vessella RL, Corey E (2010) Bone morphogenetic protein 7 is expressed in prostate cancer metastases and its effects on prostate tumor cells depend on cell phenotype and the tumor microenvironment. Neoplasia 12:192–205PubMedGoogle Scholar
  29. 29.
    Padua D, Massagué J (2009) Roles of TGFbeta in metastasis. Cell Res 19:89–102PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Hirofumi Sakai
    • 1
    • 4
  • Mutsuo Furihata
    • 2
  • Chie Matsuda
    • 1
  • Munehisa Takahashi
    • 1
  • Hiroshi Miyazaki
    • 3
  • Takeo Konakahara
    • 4
  • Toru Imamura
    • 1
  • Tomoko Okada
    • 1
  1. 1.Signaling Molecules Group, Biomedical Research InstituteNational Institute of Advanced Industrial Science and Technology (AIST)TsukubaJapan
  2. 2.Department of PathologyKochi Medical SchoolNankokuJapan
  3. 3.Virginia Commonwealth University, Philips InstituteRichmondUSA
  4. 4.Faculty of Science and TechnologyTokyo University of ScienceNodaJapan

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