Omics-based Profiling of Carcinoma of the Breast and Matched Regional Lymph Node Metastasis

Axillary lymph node (ALN) status is currently used as an important clinical indicator of breast cancer prognosis. However, the molecular mechanisms underlying lymph node metastasis are poorly understood and the relationship between ALN metastasis and the primary tumor remains unclear. In an effort to reveal structural changes in the genome and related protein responses that may drive regional metastatic progression we have analyzed matched pairs of primary breast tumors and ALN metastases both at the genomic and proteomic levels using comparative genomic hybridization (CGH) array, quantitative high‐resolution 2‐D PAGE in combination with MS, and immunohistochemistry (IHC). Array CGH revealed a remarkable similarity in genomic aberration profiles between the matched primary tumors and the ALN metastases. Quantitative profiling of 135 known proteins also revealed striking similarities in their overall expression patterns, although we observed distinct changes in the levels of individual proteins in some sample pairs. The remarkable similarities of the overall genomic and proteomic profiles between primary tumors and matched ALN metastases are taken to suggest that, in general, key biological characteristics of the primary breast tumor are maintained in the corresponding lymph node metastases. Given that the omics‐based technologies are oblivious to changes that only occur in minor cellular subsets we validated the proteomic data using IHC, which provides protein expression information with a valuable topological component. Besides confirming the omics‐derived data, the IHC analysis revealed that in two cases the ALN metastases may have been derived from a distinct minor cell subpopulation present in the primary tumor rather than from the bulk of it.

[1]  K. Alitalo,et al.  Metastasis: Lymphangiogenesis and cancer metastasis , 2002, Nature Reviews Cancer.

[2]  Takafumi Nishizaki,et al.  Genetic alterations in primary breast cancers and their metastases: Direct comparison using modified comparative genomic hybridization , 1997, Genes, chromosomes & cancer.

[3]  R. Eckhardt,et al.  DNA index and cell cycle analysis of primary breast cancer and synchronous axillary lymph node metastases , 2005, Breast Cancer Research and Treatment.

[4]  S. Singletary,et al.  Prognostic Factors in Node-Negative Breast Cancer: A Review of Studies With Sample Size More Than 200 and Follow-Up More Than 5 Years , 2002, Annals of surgery.

[5]  Wei Zhang,et al.  Differentially expressed genes between primary cancer and paired lymph node metastases predict clinical outcome of node-positive breast cancer patients , 2007, Breast Cancer Research and Treatment.

[6]  M. J. van de Vijver,et al.  Association of C‐MYC amplification with progression from the in situ to the invasive stage in C‐MYC‐amplified breast carcinomas , 2003, The Journal of pathology.

[7]  R. Moll.  Cytokeratins as markers of differentiation in the diagnosis of epithelial tumors. , 1998, Sub-cellular biochemistry.

[8]  M. Skobe,et al.  Lymphatic function, lymphangiogenesis, and cancer metastasis , 2001, Microscopy research and technique.

[9]  D. Albertson,et al.  Chromosome aberrations in solid tumors , 2003, Nature Genetics.

[10]  X. Wang,et al.  p53 tumor‐suppressor gene: Clues to molecular carcinogenesis , 1997, Journal of cellular physiology.

[11]  D. Tarin,et al.  Gene expression profiling of human lymph node metastases and matched primary breast carcinomas: Clinical implications , 2007, Molecular oncology.

[12]  M. Henriksson,et al.  Proteins of the Myc network: essential regulators of cell growth and differentiation. , 1996, Advances in cancer research.

[13]  K. Hunter.  Allelic diversity in the host genetic background may be an important determinant in tumor metastatic dissemination. , 2003, Cancer letters.

[14]  T. Dimpfl,et al.  Patients with Recurrent Breast Cancer: Does the Primary Axillary Lymph node Status Predict more Aggressive Tumor Progression? , 2003, Breast Cancer Research and Treatment.

[15]  Huanming Yang,et al.  High-resolution mapping of genotype-phenotype relationships in cri du chat syndrome using array comparative genomic hybridization. , 2005, American journal of human genetics.

[16]  R. Zeillinger,et al.  Mapping of DNA amplifications at 15 chromosomal localizations in 1875 breast tumors: definition of phenotypic groups. , 1997, Cancer research.

[17]  N. Natarajan,et al.  Breast cancer in the medial half. Results of 1978 national survey of the American College of surgeons , 1983, Cancer.

[18]  H. Beug,et al.  Molecular requirements for epithelial-mesenchymal transition during tumor progression. , 2005, Current opinion in cell biology.

[19]  J. E. Celis,et al.  Cell Biology: A Laboratory Handbook , 1997 .

[20]  P. Nowell.  The clonal evolution of tumor cell populations. , 1976, Science.

[21]  Annuska M Glas,et al.  Gene expression profiles of primary breast tumors maintained in distant metastases , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Charis Eng,et al.  Direct evidence for epithelial-mesenchymal transitions in breast cancer. , 2008, Cancer research.

[23]  Wei Zhang,et al.  Distinguishing key biological pathways between primary breast cancers and their lymph node metastases by gene function-based clustering analysis. , 2004, International journal of oncology.

[24]  S. Krishnamurthy.  Morphological and Immunophenotypic Analysis of Breast Carcinomas With Basal and Myoepithelial Differentiation , 2007 .

[25]  H. Dai,et al.  No common denominator for breast cancer lymph node metastasis , 2005, British Journal of Cancer.

[26]  Lars Bolund,et al.  Integrating Proteomic and Functional Genomic Technologies in Discovery-driven Translational Breast Cancer Research* , 2003, Molecular & Cellular Proteomics.

[27]  J. Celis,et al.  Down-regulation of the Tumor Suppressor Protein 14-3-3σ Is a Sporadic Event in Cancer of the Breast , 2005, Molecular & Cellular Proteomics.

[28]  I. Gromova,et al.  Characterization of breast precancerous lesions and myoepithelial hyperplasia in sclerosing adenosis with apocrine metaplasia , 2007, Molecular oncology.

[29]  J. Celis,et al.  Expression of the Tumor Suppressor Protein 14-3-3σ Is Down-regulated in Invasive Transitional Cell Carcinomas of the Urinary Bladder Undergoing Epithelial-to-Mesenchymal Transition* , 2004, Molecular & Cellular Proteomics.

[30]  E. Liu,et al.  Genetic background is an important determinant of metastatic potential , 2003, Nature Genetics.

[31]  B. Garcia,et al.  Proteomics , 2011, Journal of biomedicine & biotechnology.

[32]  J. Celis,et al.  High-resolution two-dimensional gel electrophoresis and protein identification using western blotting and ECL detection. , 2000, EXS.

[33]  R. Weinberg.  Is metastasis predetermined? , 2007, Molecular oncology.

[34]  J. Celis,et al.  Psoriasin (S100A7): A putative urinary marker for the follow‐up of patients with bladder squamous cell carcinomas , 1999, Electrophoresis.

[35]  K. Alitalo,et al.  Lymphangiogenesis and cancer metastasis. , 2006 .

[36]  K. Chew,et al.  Tumor labeling indices of primary breast cancers and their regional lymph node metastases , 1993, Cancer.

[37]  Satoru Miyano,et al.  Open source clustering software , 2004 .

[38]  P. Guldberg,et al.  Towards discovery‐driven translational research in breast cancer , 2004, The FEBS journal.

[39]  Jane Fridlyand,et al.  Bioinformatics Original Paper a Comparison Study: Applying Segmentation to Array Cgh Data for Downstream Analyses , 2022 .

[40]  S. Hellman,et al.  Karnofsky Memorial Lecture. Natural history of small breast cancers. , 1994, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[41]  P. O’Farrell.  High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[42]  M. Wigler,et al.  Circular binary segmentation for the analysis of array-based DNA copy number data. , 2004, Biostatistics.

[43]  J. Clements,et al.  Epithelial—mesenchymal and mesenchymal—epithelial transitions in carcinoma progression , 2007, Journal of cellular physiology.

[44]  P. Saigo,et al.  A long-term follow-up study of survival in stage I (T1N0M0) and stage II (T1N1M0) breast carcinoma. , 1989, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[45]  I. Fidler,et al.  The pathogenesis of cancer metastasis , 1980, Nature.

[46]  D. Dent,et al.  Axillary lymphadenectomy for breast cancer. Paradigm shifts and pragmatic surgeons. , 1996, Archives of surgery.

[47]  W. Woodward,et al.  Prognostic value of nodal ratios in node-positive breast cancer. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[48]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[49]  A. Shevchenko,et al.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. , 1996, Analytical chemistry.

[50]  P. Cornillet‐Lefèbvre,et al.  Compilation of published comparative genomic hybridization studies. , 2002, Cancer genetics and cytogenetics.

[51]  René Bernards,et al.  A progression puzzle. , 2002, Nature.

[52]  M G Daidone,et al.  Proliferative activity of primary breast cancer and of synchronous lymph node metastases evaluated by [3H]‐thymidine labelling index , 1990, Cell and tissue kinetics.

[53]  R. Campanini,et al.  Breast cancer metastases are molecularly distinct from their primary tumors , 2008, Oncogene.

[54]  K. Sandelin,et al.  Proteomic Characterization of the Interstitial Fluid Perfusing the Breast Tumor Microenvironment , 2004, Molecular & Cellular Proteomics.

[55]  I. Fidler,et al.  Metastasis results from preexisting variant cells within a malignant tumor. , 1977, Science.

[56]  I. Gromova,et al.  Identification of Extracellular and Intracellular Signaling Components of the Mammary Adipose Tissue and Its Interstitial Fluid in High Risk Breast Cancer Patients , 2005, Molecular & Cellular Proteomics.