Clinical & Experimental Metastasis

, Volume 27, Issue 7, pp 493–503 | Cite as

Alteration in protein expression in estrogen receptor alpha-negative human breast cancer tissues indicates a malignant and metastatic phenotype

  • Ziad J. Sahab
  • Yan-Gao Man
  • Suzan M. Semaan
  • Robert G. Newcomer
  • Stephen W. Byers
  • Qing-Xiang Amy SangEmail author
Research Paper


Ductal carcinoma in situ (DCIS) represents the earliest identifiable breast cancer lesion. Disruption of the myoepithelial cell layer and basement membrane is a prerequisite for DCIS to initiate invasion into the stroma. The majority of epithelial cells overlying a focally-disrupted myoepithelial cell layer are estrogen receptor-alpha negative (ER(−)); however, adjacent cells within the same duct confined by an intact myoepithelial cell layer express high levels of ER. These ER (+) and ER (−) cells were microdissected from the same ducts of breast cancer patients. Differential proteins expressed by ER(+) and ER(−) cells were identified using two-dimensional gel electrophoresis followed by mass spectrometry and Western blot analysis. ER(−) cells express lower levels of superoxide dismutase, RalA binding protein, galectin-1, uridine phosphorylase 2, cellular retinoic acid-binding protein 1, S100 calcium binding protein A11, and nucleoside diphosphate kinase A or non-metastasis protein 23-H1 (nm23-H1). The upregulated protein, Rho GDP-dissociation inhibitor 1 alpha, may induce chemotherapy resistance. The significant findings are that the microdissected ER(−) cells express 12.6 times less cellular retinoic acid-binding protein 1, a protein involved in cellular differentiation, and 4.1 times less nucleoside diphosphate kinase A or nm23-H1, a metastasis suppressor, and express fewer proteins than adjacent ER(+) cells. The collective role of the alterations of protein expression in ER(−) cells may be to promote a more malignant phenotype than adjacent ER(+) cells, including a decreased ability to undergo apoptosis and differentiation, and an increased potential to damage DNA, metastasize, and resist to chemotherapy.


Apoptosis Cancer heterogeneity Estrogen receptor Human breast cancer Invasion Metastasis suppressor Mass spectrometry Protein biomarker Proteomics Tumor progression 



Cellular retinoic acid-binding protein 1


Ductal carcinoma in situ


Two-dimensional gel electrophoresis




Estrogen receptor alpha


Immobilized pH gradient


Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry

ME cells

Myoepithelial cells


Molecular mass

NDPK-A (or nm23-H1)

Nucleoside diphosphate kinase A (non-metastasis protein 23-H1)




Isoelectric point


Rho GDP-dissociation inhibitor 1 alpha


S100 calcium binding protein A11


Uridine phosphorylase


Smooth muscle actin



This work was supported in part by grant BCTR0504465 from the Susan G. Komen for the Cure Breast Cancer Foundation, grants from the Florida Breast Cancer Coalition Research Foundation, the Elsa U. Pardee Foundation, and the Florida State University to Dr. Q.-X. Sang, and a grant BCTR0706983 from the Susan G. Komen Breast Cancer Foundation, grant 05AA from the AFIP/ARP joint research initiative project, and grant 2006CB910505 from the Ministry of Chinese Science and Technology to Dr. Y.-G. Man.


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Ziad J. Sahab
    • 1
    • 4
  • Yan-Gao Man
    • 2
    • 3
  • Suzan M. Semaan
    • 1
  • Robert G. Newcomer
    • 1
  • Stephen W. Byers
    • 4
  • Qing-Xiang Amy Sang
    • 1
    Email author
  1. 1.Department of Chemistry and Biochemistry and Institute of Molecular BiophysicsFlorida State UniversityTallahasseeUSA
  2. 2.Department of Gynecologic and Breast PathologyArmed Forces Institute of Pathology and American Registry of PathologyWashingtonUSA
  3. 3.Jilin UniversityChangchunChina
  4. 4.Department of Oncology, Lombardi Comprehensive Cancer CenterGeorgetown University Medical CenterWashingtonUSA

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