Clinical & Experimental Metastasis

, Volume 29, Issue 4, pp 315–325 | Cite as

Ubiquitous Brms1 expression is critical for mammary carcinoma metastasis suppression via promotion of apoptosis

  • Leah M. Cook
  • Xuemei Cao
  • Alexander E. Dowell
  • Michael T. Debies
  • Mick D. Edmonds
  • Benjamin H. Beck
  • Robert A. Kesterson
  • Renee A. Desmond
  • Andra R. Frost
  • Douglas R. HurstEmail author
  • Danny R. WelchEmail author
Research Paper


Morbidity and mortality of breast cancer patients are drastically increased when primary tumor cells are able to spread to distant sites and proliferate to become secondary lesions. Effective treatment of metastatic disease has been limited; therefore, an increased molecular understanding to identify biomarkers and therapeutic targets is needed. Breast cancer metastasis suppressor 1 (BRMS1) suppresses development of pulmonary metastases when expressed in a variety of cancer types, including metastatic mammary carcinoma. Little is known of Brms1 function throughout the initiation and progression of mammary carcinoma. The goal of this study was to investigate mechanisms of Brms1-mediated metastasis suppression in transgenic mice that express Brms1 using polyoma middle T oncogene-induced models. Brms1 expression did not significantly alter growth of the primary tumors. When expressed ubiquitously using a β-actin promoter, Brms1 suppressed pulmonary metastasis and promoted apoptosis of tumor cells located in the lungs but not in the mammary glands. Surprisingly, selective expression of Brms1 in the mammary gland using the MMTV promoter did not significantly block metastasis nor did it promote apoptosis in the mammary glands or lung, despite MMTV-induced expression within the lungs. These results strongly suggest that cell type-specific over-expression of Brms1 is important for Brms1-mediated metastasis suppression.


Metastasis BRMS1 PyMT Transgenic MMTV Ubiquitous Apoptosis Breast cancer Mouse Mammary Tumor microenvironment 



Breast cancer metastasis suppressor 1


Polyoma middle T antigen


Mouse mammary tumor virus








Terminal deoxynucleotidyl transferase dUTP nick end labeling


Terminal deoxynucleotidyl transferase


Nuclear factor kappa B


Histone deacetylase







We would like to thank Dr. Clinton Grubbs in the UAB Department of Pharmacology and Toxicology for processing and preparing all histology slides. We are grateful for the counsel of Drs. Terri Wood and Rosa Serra during the course of these studies. We also thank Dr. Monica Richert for aiding in early colony maintenance of our transgenic mice and members of the Welch and Hurst labs for critical reading of the manuscript. We apologize to those whose work could not be cited due to space limitations. This paper is published in partial fulfillment of the requirements of the doctoral dissertation of LMC. Grant Support: National Institutes of Health [CA87728 (DRW), CA134981 (DRW) and CA089019 (DRW & DRH], National Foundation for Cancer Research (DRW), and Susan G. Komen for the Cure [SAC11037] (DRW); UAB Cancer Prevention and Control Training Grant [CA47888] (LMC); American Cancer Society [RSG-11-259-01-CSM] (DRH). DRW is an Eminent Scholar of the Kansas Biosciences Authority. The Transgenic Mouse Facility and Biostatistics Shared Resources were supported by the UAB Comprehensive Cancer Center core grant P30 CA13148.

Conflict of interest

We certify there are no conflicts of interest.

Supplementary material

10585_2012_9452_MOESM1_ESM.tif (24.1 mb)
Supplemental Fig. 1 Lung tissues do not exhibit alterations in morphology or pathology of metastatic lesions. Lung specimens from Brms1 transgenic mice and control littermates were collected, FFPE, and 5μm sections were stained with H&E. There were no visible differences in lung histomorphology due to Brms1 overexpression. There were no visible differences in the pathology of metastatic lesions in Brms1Ubqs mice. All images were captured at 200x magnification, size bar = 50 μm (TIFF 24672 kb)
10585_2012_9452_MOESM2_ESM.tif (24.1 mb)
Supplemental Fig. 2 Organ tissues do not exhibit metastatic development or alterations in morphology. Tissue specimens from Brms1 transgenic mice and control littermates were collected, FFPE, and 5μm sections were stained with H&E. There were no visible differences in organ histomorphology due to Brms1 overexpression. There were no microscopic metastases detected. All images were captured at 200x magnification, size bar = 50 μm (TIFF 24670 kb)


  1. 1.
    Siegel R, Ward E, Brawley O et al (2011) Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin 61:212–236PubMedCrossRefGoogle Scholar
  2. 2.
    Cook LM, Hurst DR, Welch DR (2011) Metastasis suppressors and the tumor microenvironment. Semin Cancer Biol 21:113–122PubMedCrossRefGoogle Scholar
  3. 3.
    Hurst DR, Welch DR (2011) Metastasis suppressor genes: at the interface between the environment and tumor cell growth. Int Rev Cell Mol Biol 286:107–180PubMedCrossRefGoogle Scholar
  4. 4.
    Seraj MJ, Samant RS, Verderame MF et al (2000) Functional evidence for a novel human breast carcinoma metastasis suppressor, BRMS1, encoded at chromosome 11q13. Cancer Res 60:2764–2769PubMedGoogle Scholar
  5. 5.
    Shevde LA, Samant RS, Goldberg SF et al (2002) Suppression of human melanoma metastasis by the metastasis suppressor gene, BRMS1. Exp Cell Res 273:229–239PubMedCrossRefGoogle Scholar
  6. 6.
    Zhang S, Lin QD, Di W (2006) Suppression of human ovarian carcinoma metastasis by the metastasis-suppressor gene, BRMS1. Int J Gynecol Cancer 16:522–531PubMedCrossRefGoogle Scholar
  7. 7.
    Smith PW, Liu Y, Siefert SA et al (2009) Breast cancer metastasis suppressor 1 (BRMS1) suppresses metastasis and correlates with improved patient survival in non-small cell lung cancer. Cancer Lett 276:196–203PubMedCrossRefGoogle Scholar
  8. 8.
    Saunders MM, Seraj MJ, Li ZY et al (2001) Breast cancer metastatic potential correlates with a breakdown in homospecific and heterospecific gap junctional intercellular communication. Cancer Res 61:1765–1767PubMedGoogle Scholar
  9. 9.
    Kapoor P, Saunders MM, Li Z et al (2004) Breast cancer metastatic potential: correlation with increased heterotypic gap junctional intercellular communication between breast cancer cells and osteoblastic cells. Int J Cancer 111:693–697PubMedCrossRefGoogle Scholar
  10. 10.
    Bodenstine TM, Vaidya KS, Ismail A et al (2010) Homotypic gap junctional communication associated with metastasis suppression increases with PKA activity and is unaffected by PI3K inhibition. Cancer Res 70:10002–10011PubMedCrossRefGoogle Scholar
  11. 11.
    DeWald DB, Torabinejad J, Samant RS et al (2005) Metastasis suppression by breast cancer metastasis suppressor 1 involves reduction of phosphoinositide signaling in MDA-MB-435 breast carcinoma cells. Cancer Res 65:713–717PubMedGoogle Scholar
  12. 12.
    Vaidya KS, Harihar S, Stafford LJ et al (2008) Breast cancer metastasis suppressor-1 differentially modulates growth factor signaling. J Biol Chem 283:28354–28360PubMedCrossRefGoogle Scholar
  13. 13.
    Cicek M, Fukuyama R, Welch DR et al (2005) Breast cancer metastasis suppressor 1 inhibits gene expression by targeting nuclear factor-kappaB activity. Cancer Res 65:3586–3595PubMedCrossRefGoogle Scholar
  14. 14.
    Samant RS, Clark DW, Fillmore RA et al (2007) Breast cancer metastasis suppressor 1 (BRMS1) inhibits osteopontin transcription by abrogating NF-kappaB activation. Mol Cancer 6:6PubMedCrossRefGoogle Scholar
  15. 15.
    Liu Y, Smith PW, Jones DR (2006) Breast cancer metastasis suppressor 1 functions as a corepressor by enhancing histone deacetylase 1-mediated deacetylation of RelA/p65 and promoting apoptosis. Mol Cell Biol 26:8683–8696PubMedCrossRefGoogle Scholar
  16. 16.
    Samant RS, Seraj MJ, Saunders MM et al (2001) Analysis of mechanisms underlying BRMS1 suppression of metastasis. Clin Exp Metastasis 18:683–693CrossRefGoogle Scholar
  17. 17.
    Phadke PA, Vaidya KS, Nash KT et al (2008) BRMS1 suppresses breast cancer experimental metastasis to multiple organs by inhibiting several steps of the metastatic process. Am J Pathol 172:809–817PubMedCrossRefGoogle Scholar
  18. 18.
    Meehan WJ, Samant RS, Hopper JE et al (2004) Breast cancer metastasis suppressor 1 (BRMS1) forms complexes with retinoblastoma-binding protein 1 (RBP1) and the mSin3 histone deacetylase complex and represses transcription. J Biol Chem 279:1562–1569PubMedCrossRefGoogle Scholar
  19. 19.
    Hurst DR, Xie Y, Vaidya KS et al (2008) Alterations of BRMS1–ARID4A interaction modify gene expression but still suppress metastasis in human breast cancer cells. J Biol Chem 283:7438–7444PubMedCrossRefGoogle Scholar
  20. 20.
    Cicek M, Fukuyama R, Cicek MS et al (2009) BRMS1 contributes to the negative regulation of uPA gene expression through recruitment of HDAC1 to the NF-kappaB binding site of the uPA promoter. Clin Exp Metastasis 26:229–237PubMedCrossRefGoogle Scholar
  21. 21.
    Samant RS, Debies MT, Shevde LA et al (2002) Identification and characterization of murine ortholog (Brms1) of breast cancer metastasis suppressor 1 (BRMS1). Int J Cancer 97:15–20PubMedCrossRefGoogle Scholar
  22. 22.
    Samant RS, Debies MT, Hurst DR et al (2006) Suppression of murine mammary carcinoma metastasis by the murine ortholog of breast cancer metastasis suppressor 1 (Brms1). Cancer Lett 235:260–265PubMedCrossRefGoogle Scholar
  23. 23.
    Richmond A, Su Y (2008) Mouse xenograft models vs. GEM models for human cancer therapeutics. Dis Model Mech 1:78–82PubMedCrossRefGoogle Scholar
  24. 24.
    Welch DR (1997) Technical considerations for studying cancer metastasis in vivo. Clin Exp Metastasis 15:272–306PubMedCrossRefGoogle Scholar
  25. 25.
    Guy CT, Cardiff RD, Muller WJ (1992) Induction of mammary tumors by expression of polyomavirus middle T oncogene: a transgenic mouse model for metastatic disease. Mol Cell Biol 12:954–961PubMedGoogle Scholar
  26. 26.
    Lin EY, Jones JG, Li P et al (2003) Progression to malignancy in the polyoma middle T oncoprotein mouse breast cancer model provides a reliable model for human diseases. Am J Pathol 163:2113–2126PubMedCrossRefGoogle Scholar
  27. 27.
    Hurst DR, Edmonds MD, Scott GK et al (2009) Breast cancer metastasis suppressor 1 BRMS1 up-regulates miR-146 that suppresses breast cancer metastasis. Cancer Res 69:1279–1283PubMedCrossRefGoogle Scholar
  28. 28.
    Weidner N (1995) Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors. Breast Cancer Res Treat 36:169–180PubMedCrossRefGoogle Scholar
  29. 29.
    Henrard D, Ross SR (1988) Endogenous mouse mammary tumor virus is expressed in several organs in addition to the lactating mammary gland. J Virol 62:3046–3049PubMedGoogle Scholar
  30. 30.
    Schneider J, Gomez-Esquer F, Diaz-Gil G et al (2011) mRNA expression of the putative antimetastatic gene BRMS1 and of apoptosis-related genes in breast cancer. Cancer Genomics Proteomics 8:195–197PubMedGoogle Scholar
  31. 31.
    Ladeda V, Adam AP, Puricelli L et al (2001) Apoptotic cell death in mammary adenocarcinoma cells is prevented by soluble factors present in the target organ of metastasis. Breast Cancer Res Treat 69:39–51PubMedCrossRefGoogle Scholar
  32. 32.
    Cavanaugh PG, Nicolson GL (1989) Purification and some properties of a lung-derived growth factor that differentially stimulates the growth of tumor cells metastatic to the lung. Cancer Res 49:3928–3933PubMedGoogle Scholar
  33. 33.
    Wyckoff JB, Wang Y, Lin EY et al (2007) Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res 67:2649–2656PubMedCrossRefGoogle Scholar
  34. 34.
    Wang W, Wyckoff JB, Goswami S et al (2007) Coordinated regulation of pathways for enhanced cell motility and chemotaxis is conserved in rat and mouse mammary tumors. Cancer Res 67:3505–3511PubMedCrossRefGoogle Scholar
  35. 35.
    Grum-Schwensen B, Klingelhofer J, Grigorian M et al (2010) Lung metastasis fails in MMTV-PyMT oncomice lacking S100A4 due to a T-cell deficiency in primary tumors. Cancer Res 70:936–947PubMedCrossRefGoogle Scholar
  36. 36.
    DeNardo DG, Barreto JB, Andreu P et al (2009) CD4(+) T cells regulate pulmonary metastasis of mammary carcinomas by enhancing protumor properties of macrophages. Cancer Cell 16:91–102PubMedCrossRefGoogle Scholar
  37. 37.
    Yang L, Huang J, Ren X et al (2008) Abrogation of TGF beta signaling in mammary carcinomas recruits Gr-1+CD11b+myeloid cells that promote metastasis. Cancer Cell 13:23–35PubMedCrossRefGoogle Scholar
  38. 38.
    Wiseman BS, Werb Z (2002) Stromal effects on mammary gland development and breast cancer. Science (Washington, DC) 296:1046–1049CrossRefGoogle Scholar
  39. 39.
    Bhowmick NA, Neilson EG, Moses HL (2004) Stromal fibroblasts in cancer initiation and progression. Nature 432:332–337PubMedCrossRefGoogle Scholar
  40. 40.
    Lifsted T, Le Voyer T, Williams M et al (1998) Identification of inbred mouse strains harboring genetic modifiers of mammary tumor age of onset and metastatic progression. Int J Cancer 77:640–644PubMedCrossRefGoogle Scholar
  41. 41.
    Qiu TH, Chandramouli GVR, Hunter KW et al (2004) Global expression profiling identifies signatures of tumor virulence in MMTV-PyMT-transgenic mice: correlation to human disease. Cancer Res 64:5973–5981PubMedCrossRefGoogle Scholar
  42. 42.
    Zhang Z, Yamashita H, Toyama T et al (2006) Reduced expression of the breast cancer metastasis suppressor 1 mRNA is correlated with poor progress in breast cancer. Clin Cancer Res 12:6410–6414PubMedCrossRefGoogle Scholar
  43. 43.
    Hicks DG, Yoder BJ, Short S et al (2006) Loss of BRMS1 protein expression predicts reduced disease-free survival in hormone receptor negative and HER2 positive subsets of breast cancer. Clin Cancer Res 12:6702–6708PubMedCrossRefGoogle Scholar
  44. 44.
    Lombardi G, Di Cristofano C, Capodanno A et al (2006) High level of messenger RNA for BRMS1 in primary breast carcinomas is associated with poor prognosis. Int J Cancer 120:1169–1178CrossRefGoogle Scholar
  45. 45.
    Metge BJ, Frost AR, King JA et al (2008) Epigenetic silencing contributes to the loss of BRMS1 expression in breast cancer. Clin Exp Metastasis 25:753–763PubMedCrossRefGoogle Scholar
  46. 46.
    Frolova N, Edmonds MD, Bodenstine TM et al (2009) A shift from nuclear to cytoplasmic breast cancer metastasis suppressor 1 expression is associated with highly proliferative estrogen receptor-negative breast cancers. Tumour Biol 30:148–159PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Leah M. Cook
    • 1
  • Xuemei Cao
    • 1
    • 7
  • Alexander E. Dowell
    • 2
  • Michael T. Debies
    • 7
  • Mick D. Edmonds
    • 1
  • Benjamin H. Beck
    • 1
  • Robert A. Kesterson
    • 3
    • 6
  • Renee A. Desmond
    • 2
    • 6
  • Andra R. Frost
    • 1
    • 4
    • 6
  • Douglas R. Hurst
    • 1
    • 6
    Email author
  • Danny R. Welch
    • 1
    • 4
    • 5
    • 6
    • 7
    • 8
    Email author
  1. 1.Department of PathologyUniversity of Alabama at BirminghamBirminghamUSA
  2. 2.Department of MedicineUniversity of Alabama at BirminghamBirminghamUSA
  3. 3.Department of GeneticsUniversity of Alabama at BirminghamBirminghamUSA
  4. 4.Department of Cell BiologyUniversity of Alabama at BirminghamBirminghamUSA
  5. 5.Department of Pharmacology and ToxicologyUniversity of Alabama at BirminghamBirminghamUSA
  6. 6.Comprehensive Cancer CenterUniversity of Alabama at BirminghamBirminghamUSA
  7. 7.Department of Cancer BiologyThe Kansas University Medical CenterKansas CityUSA
  8. 8.University of Kansas Cancer CenterThe Kansas University Medical CenterKansas CityUSA

Personalised recommendations