Clinical & Experimental Metastasis

, Volume 21, Issue 5, pp 427–435 | Cite as

Metastatic breast cancer cells suppress osteoblast adhesion and differentiation

  • Robyn R. Mercer
  • Chiaki Miyasaka
  • Andrea M. MastroEmail author


Bone is a primary target for colonization of metastatic breast cancer cells. Once present, the breast cancer cells activate osteoclasts, thereby stimulating bone loss. Bone degradation is accompanied by pain and increased susceptibility to fractures. However, targeted inhibition of osteoclasts does not completely prevent lesion progression, nor does it heal the lesions. This suggests that breast cancer cells may also affect osteoblasts, cells that build bone. The focus of this study was to determine the ability of breast cancer cells to alter osteoblast function. MC3T3-E1 osteoblasts were cultured with conditioned medium from MDA-MB-231 breast cancer cells and subsequently assayed for changes in differentiation. Osteoblast differentiation was monitored by expression of osteocalcin, bone sialoprotein and alkaline phosphatase, and by mineralization. Osteoblasts cultured with MDA-MB-231 conditioned medium did not express these mature bone proteins, nor did they mineralize a matrix. Inhibition of osteoblast differentiation was found to be due to transforming growth factor β present in MDA-MB-231 conditioned medium. Interestingly, breast cancer conditioned medium also altered cell adhesion. When osteoblasts were assayed for adhesion properties using interference reflection microscopy and scanning acoustic microscopy, there was a reduction in focal adhesion plaques and sites of detachment were clearly visible. F-actin was disassembled and punctate in osteoblasts cultured with MDA-MB-231 conditioned medium rather than organized in long stress fibers. Taken together, these observations suggest that metastatic breast cancer cells alter osteoblast adhesion and prevent differentiation. These affects could account for the continued loss of bone after osteoclast inhibition in patients with bone-metastatic breast cancer.


adhesion bone breast cancer differentiation metastasis osteoblasts 



bone sialoprotein


interference reflection microscopy






transforming growth factor β


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Landis, SH, Murray, T, Bolden, S,  et al. 1999Cancer statisticsClinical & Experimental Metastasis49831Google Scholar
  2. Galasko, CS 1982Mechanisms of lytic and blastic metastatic disease of boneClinical & Experimental Metastasis169207Google Scholar
  3. Guise, TA, Mundy, GR 1998Cancer and boneClinical & Experimental Metastasis191854Google Scholar
  4. Stewart, AF, Vignery, A, Silverglate, A,  et al. 1982Quantitative bone histomorphometry in humoral hypercalcemia of malignancy: uncoupling of bone cell activityClinical & Experimental Metastasis5521927Google Scholar
  5. Kukreja, SC, Rosol, TJ, Shevrin, DH,  et al. 1988Quantitative bone histomorphometry in nude mice bearing a human squamous cell lung cancerClinical & Experimental Metastasis33416Google Scholar
  6. Delmas, PD, Demiaux, B, Malaval, L,  et al. 1986Serum bone gamma carboxyglutamic acid-containing protein in primary hyperparathyroidism and in malignant hypercalcemia. Comparison with bone histomorphometryClinical & Experimental Metastasis7798591Google Scholar
  7. Taube, T, Elomaa, I, Blomqvist, C,  et al. 1994Histomorphometric evidence for osteoclast-mediated bone resorption in metastatic breast cancerClinical & Experimental Metastasis151616Google Scholar
  8. Mastro, AM, Gay, CV, Welch, DR,  et al. 2004Breast cancer cells induce osteoblast apoptosis: A possible contributor to bone degradationClinical & Experimental Metastasis9126576Google Scholar
  9. Fromigue, O, Kheddoumi, N, Lomri, A,  et al. 2001Breast cancer cells release factors that induced apoptosis in human bone marrow stromal cellsClinical & Experimental Metastasis16160010Google Scholar
  10. Lacroix, M, Siwek, B, Body, JJ 1996Effects of secretory products of breast cancer cells on osteoblast-like cellsClinical & Experimental Metastasis3820916Google Scholar
  11. Siwek, B, Lacroix, M, De Pollak, C,  et al. 1997Secretory products of breast cancer cells specifically affect human osteoblastic cells: partial characterization of active factorsClinical & Experimental Metastasis1255260Google Scholar
  12. Bunyaratavej, P, Hullinger, TG, Somerman, MJ 2000Bone morphogenetic proteins secreted by breast cancer cells upregulate bone sialoprotein expression in preosteoblast cellsClinical & Experimental Metastasis26032433Google Scholar
  13. Hullinger, TG, Taichman, RS, Linseman, DA,  et al. 2000Secretory products from PC-3 and MCF-7 tumor cell lines upregulate osteopontin in MC3T3-E1 cellsClinical & Experimental Metastasis7860716Google Scholar
  14. Quarles, LD, Yohay, DA, Lever, LW,  et al. 1992Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast developmentClinical & Experimental Metastasis768392Google Scholar
  15. Cailleau, R, Young, R, Olive, M,  et al. 1974Breast tumor cell lines from pleural effusionsClinical & Experimental Metastasis5366174Google Scholar
  16. Yoneda, T, Sasaki, A, Mundy, GR 1994Osteolytic bone metastasis in breast cancerClinical & Experimental Metastasis327384Google Scholar
  17. Hildebrand, JA, Rugar, D, Johnston, RN,  et al. 1981Acoustic microscopy of living cellsClinical & Experimental Metastasis78165660Google Scholar
  18. Lian, JB, Stein, GS 1992Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formationClinical & Experimental Metastasis3269305Google Scholar
  19. Pederson, L, Winding, B, Foged, NT,  et al. 1999Identification of breast cancer cell line-derived paracrine factors that stimulate osteoclast activityClinical & Experimental Metastasis59584955Google Scholar
  20. Bonewald LF. Transforming growth factor-β. In Principles of Bone Biology, 2nd ed, Vol. 2. San Diego: Academic Press 2002; 903–18.Google Scholar
  21. Spinella-Jaegle, S, Roman-Roman, S, Faucheu, C,  et al. 2001Opposite effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on osteoblast differentiationClinical & Experimental Metastasis2932330Google Scholar
  22. Damsky, CH 1999Extracellular matrix-integrin interactions in osteoblast function and tissue remodelingClinical & Experimental Metastasis25956Google Scholar
  23. Bennett, JH, Moffatt, S, Horton, M 2001Cell adhesion molecules in human osteoblasts: structure and functionClinical & Experimental Metastasis1660311Google Scholar
  24. Xiao, G, Wang, D, Benson, MD,  et al. 1998Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factorClinical & Experimental Metastasis2733298894Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Robyn R. Mercer
    • 1
    • 3
  • Chiaki Miyasaka
    • 2
  • Andrea M. Mastro
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of Biochemistry and Molecular Biologythe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.Department of Engineering Sciences and Mechanicsthe Pennsylvania State UniversityUniversity ParkUSA
  3. 3.National Foundation for Cancer Research, Center for Metastasis ResearchUniversity of Alabama at BirminghamBirminghamUSA
  4. 4.Andrea MastroUniversity ParkUSA

Personalised recommendations