Genomic Alterations Associated with Early Stages of Breast Tumor Metastasis

  • Rachel E. Ellsworth
  • Darrell L. Ellsworth
  • Heather L. Patney
  • Brenda Deyarmin
  • Jeffrey A. Hooke
  • Brad Love
  • Craig D. Shriver
Breast Oncology



Molecular studies suggest that acquisition of metastatic potential occurs early in the development of breast cancer; mechanisms by which cells disseminate from the primary carcinomas and successfully colonize foreign tissues are, however, largely unknown. Thus, we examined levels and patterns of chromosomal alterations in primary breast tumors from node-negative (n = 114) and node-positive (n = 115) patients to determine whether specific genomic changes are associated with tumor metastasis.


Fifty-two genetic markers representing 26 chromosomal regions commonly altered in breast cancer were examined in laser microdissected tumor samples to assess levels and patterns of allelic imbalance (AI). Real time-PCR (RT-PCR) was performed to determine expression levels of candidate genes. Data was analyzed using exact unconditional and Student’s t-tests with significance values of P < 0.05 and P < 0.002 used for the clinicopathological and genomic analyses, respectively.


Overall levels of AI in primary breast tumors from node-negative (20.8%) and node-positive (21.9%) patients did not differ significantly (P = 0.291). When data were examined by chromosomal region, only chromosome 8q24 showed significantly higher levels (P < 0.0005) of AI in node-positive primary tumors (23%) versus node-negative samples (6%). c-MYC showed significantly higher levels of gene expression in primary breast tumors from patients with lymph node metastasis.


Higher frequencies of AI at chromosome 8q24 in patients with positive lymph nodes suggest that genetic changes in this region are important to the process of metastasis. Because overexpression of c-MYC has been associated with cellular dissemination as well as development of the premetastatic niche, alterations of the 8q24 region, including c-MYC, may be key determinants in the development of lymph node metastasis.


Allelic imbalance Primary breast tumor Metastasis 



Presented in part at the Society of Surgical Oncology’s 61st Annual Cancer Symposium, March 13—16, 2008 in Chicago, IL. This work was performed under the auspices of the Clinical Breast Care Project, a joint effort of many investigators and staff members whose contributions are gratefully acknowledged. We especially thank the program participants. Supported by the United States Department of Defense (Military Molecular Medicine Initiative MDA W81XWH-05–2–0075). The opinion and assertions contained herein are the private views of the authors and are not to be construed as official or as representing the views of the Department of the Army or the Department of Defense.


  1. 1.
    Eifel P, Axelson JA, Costa J, et al. National Institutes of Health Consensus Development Conference Statement: adjuvant therapy for breast cancer, November 1–3, 2000. J Natl Cancer Inst 2001;93:979–89PubMedCrossRefGoogle Scholar
  2. 2.
    National Cancer Institute. SEER cancer statistics review, 1973–1991. NIH Publ. No. 94–2789. 1994. Bethesda, MD, NIHGoogle Scholar
  3. 3.
    Ellsworth RE, Ellsworth DL, Neatrour DM, et al. Allelic imbalance in primary breast carcinomas and metastatic tumors of the axillary lymph nodes. Mol Cancer Res 2005;3:71–7PubMedCrossRefGoogle Scholar
  4. 4.
    Kuukasjarvi T, Karhu R, Tanner M, et al. Genetic heterogeneity and clonal evolution underlying development of asynchronous metastasis in human breast cancer. Cancer Res 1997; 57:1597–604PubMedGoogle Scholar
  5. 5.
    Schmidt-Kittler O, Ragg T, Daskalakis A, et al. From latent disseminated cells to overt metastasis: genetic analysis of systemic breast cancer progression. Proc Natl Acad Sci USA 2003; 100:7737–42PubMedCrossRefGoogle Scholar
  6. 6.
    Newman B, Austin MA, Lee M, et al. Inheritance of human breast cancer: evidence for autosomal dominant transmission in high-risk families. Proc Natl Acad Sci 1988; 85:3044–8PubMedCrossRefGoogle Scholar
  7. 7.
    Page DL, Dupont WD, Rogers LW, et al. Intraductal carcinoma of the breast: follow-up after biopsy only. Cancer 1982; 49:751–8PubMedCrossRefGoogle Scholar
  8. 8.
    Ellsworth DL, Shriver CD, Ellsworth RE, et al. Laser capture microdissection of paraffin-embedded tissues. BioTechniques 2003; 34:42–6PubMedGoogle Scholar
  9. 9.
    Ellsworth RE, Ellsworth DL, Lubert SM, et al. High-throughput loss of heterozygosity mapping in 26 commonly deleted regions in breast cancer. Cancer Epidemiol Biomarkers Prev 2003; 12:915–9PubMedGoogle Scholar
  10. 10.
    Medintz IL, Lee C-CR, Wong WW, et al. Loss of heterozygosity assay for molecular detection of cancer using energy-transfer primers and capillary array electrophoresis. Genome Res 2000; 10:1211–8PubMedCrossRefGoogle Scholar
  11. 11.
    Sleeman JP. The lymph node as a bridgehead in the metastatic dissemination of tumors. Recent Results Cancer Res 2000; 157:55–81PubMedGoogle Scholar
  12. 12.
    Hartveit E. Attenuated cells in breast stroma: the missing lymphatic system of the breast. Histopathology 1990; 16:533–43PubMedCrossRefGoogle Scholar
  13. 13.
    Kaplan RN, Riba RD, Zacharoulis S, et al. VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche. Nature 2005; 438:820–7PubMedCrossRefGoogle Scholar
  14. 14.
    Hiratsuka S, Watanabe A, Aburatani H, et al. Tumour-mediated upregulation of chemoattractants and recruitment of myeloid cells predetermines lung metastasis. Nat Cell Biol 2006; 8:1369–75PubMedCrossRefGoogle Scholar
  15. 15.
    Fearon ER, Vogelstein B. A genetic model for colorectal tumorigenesis. Cell 1990; 61:759–67PubMedCrossRefGoogle Scholar
  16. 16.
    Bissig H, Richter J, Desper R, et al. Evaluation of the clonal relationship between primary and metastatic renal cell carcinoma by comparative genomic hybridization. Am J Pathol 1999; 155:267–74PubMedGoogle Scholar
  17. 17.
    Sasatomi E, Finkelstein SD, Woods JD, et al. Comparison of accumulated allele loss between primary tumor and lymph node metastasis in stage II non-small cell lung carcinoma: implications for the timing of lymph node metastasis and prognostic value. Cancer Res 2002; 62:2681–9PubMedGoogle Scholar
  18. 18.
    Kerbel RS. Growth dominance of the metastatic cancer cell: cellular and molecular aspects. Adv Cancer Res 1990; 55:87–132PubMedCrossRefGoogle Scholar
  19. 19.
    Nowell PC. The clonal evolution of tumor cell populations. Science 1976; 194:23–8PubMedCrossRefGoogle Scholar
  20. 20.
    Poste G, Fidler IJ. The pathogenesis of cancer metastasis. Nature 1980; 283:139–46PubMedCrossRefGoogle Scholar
  21. 21.
    Ramaswamy S, Ross KN, Lander ES, et al. A molecular signature of metastasis in primary solid tumors. Nat Genet 2003; 33:49–54PubMedCrossRefGoogle Scholar
  22. 22.
    van’t Veer LJ, Dai H, vande Vijver MJ, et al. Gene expression profiling predicts clincial outcome of breast cancer. Nature 2002; 415:530–6CrossRefGoogle Scholar
  23. 23.
    van de Vijer MJ, He YD, van’t Veer LJ, et al. A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med 2002; 347:1999–2009CrossRefGoogle Scholar
  24. 24.
    Weigelt B, Glas AM, Wessels LF, et al. Gene expression profiles of primary breast tumors maintained in distant metastases. Proc Natl Acad Sci USA 2003; 100:15901–5PubMedCrossRefGoogle Scholar
  25. 25.
    Sleeman JP, Cremers N. New concepts in breast cancer metastasis: tumor initiating cells and the microenvironment. Clin Exp Metastasis 2007; 24:707–15PubMedCrossRefGoogle Scholar
  26. 26.
    Kang Y, Siegel PM, Shu W, et al. A multigenic program mediating breast cancer metastasis to bone. Cancer Cell 2003; 3:537–49PubMedCrossRefGoogle Scholar
  27. 27.
    Minn AJ, Kang Y, Serganova I, et al. Distinct organ-specific metastatic potential of individual breast cancer cells and primary tumors. J Clin Invest 2005; 115:44–55PubMedGoogle Scholar
  28. 28.
    Kerangueven F, Essioux L, Dib A, et al. Loss of heterozygosity and linkage analysis in breast carcinoma: indication for a putative third susceptibility gene on the short arm of chromosome 8. Oncogene 1995; 10:1023–6PubMedGoogle Scholar
  29. 29.
    Lichy JH, Dalbegue F, Zavar M, et al. Genetic heterogeneity in ductal carcinoma of the breast. Lab Invest 2000; 80:291–301PubMedGoogle Scholar
  30. 30.
    Hermsen MA, Baak JP, Meijer GA, et al. Genetic analysis of 53 lymph node-negative breast carcinomas by CGH and relation to clinical, pathological, morphometric, and DNA cytometric prognostic factors. J Pathol 1998; 186:356–62PubMedCrossRefGoogle Scholar
  31. 31.
    Guerin M, Barrois M, Terrier MJ, et al. Overexpression of either c-myc or c-erB-2/neu proto-oncogenes in human breast carcinomas: correlation with poor prognosis. Oncogene Res 1988; 3:21–31PubMedGoogle Scholar
  32. 32.
    Varley JM, Swallow JE, Brammar WJ, et al. Alterations to either c-erbB(neu) or c-myc proto-congoenes in breast carcinomas correlate with poor short-term prognosis. Oncogene 1987; 1:423–30PubMedGoogle Scholar
  33. 33.
    Machotka SV, Garrett CT, Schwartz AM, et al. Amplification of the proto-oncogenes int-2, c-erb B-2 and c-myc in human breast cancer. Clin Chim Acta 1989; 184:207–17PubMedCrossRefGoogle Scholar
  34. 34.
    Nagayama K, Watatani M. Analysis of genetic alterations related to the development and progression of breast carcinoma. Jpn J Cancer Res 1993; 84:1159–64PubMedGoogle Scholar
  35. 35.
    Levens D. Disentangling the MYC web. Proc Natl Acad Sci USA 2002; 99:5757–9PubMedCrossRefGoogle Scholar
  36. 36.
    Brouillet JP, Theillet C, Maudelonde T, et al. Cathepsin D assay in primary breast cancer and lymph nodes: relationship with c-myc, c-erb-B-2 and int-2 oncogene amplification and node invasiveness. Eur J Cancer 1990; 26:437–41PubMedCrossRefGoogle Scholar
  37. 37.
    Giambernardi TA, Grant GM, Taylor GP, et al. Overview of matrix metalloproteinase expression in cultured human cells. Matrix Biol 1998; 16:483–96PubMedCrossRefGoogle Scholar
  38. 38.
    Kaplan RN, Rafii S, Lyden D. Preparing the “Soil”: the premetastatic niche. Cancer Res 2006; 66:11089–93PubMedCrossRefGoogle Scholar
  39. 39.
    Baudino TA, McKay C, Pendeville-Samain H, et al. c-Myc is essential for vasculogenesis and angiogenesis during development and tumor progression. Genes Dev 2002; 16:2530–43PubMedCrossRefGoogle Scholar

Copyright information

© Society of Surgical Oncology 2008

Authors and Affiliations

  • Rachel E. Ellsworth
    • 1
    • 2
  • Darrell L. Ellsworth
    • 2
  • Heather L. Patney
    • 2
  • Brenda Deyarmin
    • 2
  • Jeffrey A. Hooke
    • 3
  • Brad Love
    • 4
  • Craig D. Shriver
    • 3
  1. 1.Clinical Breast Care ProjectHenry M. Jackson Foundation for the Advancement of Military MedicineWindberUSA
  2. 2.Clinical Breast Care ProjectWindber Research InstituteWindberUSA
  3. 3.Clinical Breast Care ProjectWalter Reed Army Medical CenterWashingtonUSA
  4. 4.InvitrogenCarlsbadUSA

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