J Breast Cancer.  2015 Sep;18(3):242-248. 10.4048/jbc.2015.18.3.242.

The Presence of EpCAM-/CD49f+ Cells in Breast Cancer Is Associated with a Poor Clinical Outcome

• Ye F
• Qiu Y
• Li L
• Yang L
• Cheng F
• Zhang H
• Wei B
• Zhang Z
• Sun L
• Bu H

• KMID: 2407571
• DOI: http://doi.org/10.4048/jbc.2015.18.3.242

Abstract

PURPOSE
It is well established that breast cancer stem cells (BCSCs) play an essential role in tumor invasion for both local and distant metastasis. The aim of this study was to establish whether BCSCs could act as a prognostic and clinical marker.
METHODS
We analyzed tumor tissues from 161 breast cancer patients. Dual immunohistochemistry and immunofluorescence were performed on all the slides, and we analyzed the relationship between EpCAM-/CD49f+ tumor cells and key clinical and prognostic factors.
RESULTS
Univariate survival analysis using the Kaplan-Meier method showed that the presence of EpCAM-/CD49f+ tumor cells in breast cancer was significantly associated with shorter disease-free survival (DFS) and overall survival (OS). Multivariate analysis showed that the presence of EpCAM-/CD49f+ cells was associated with shorter DFS (p=0.010; hazard ratio [HR], 2.070) and OS (p=0.002; HR, 3.235). Tumors containing EpCAM-/CD49f+ cells were also more likely to metastasize after initial surgery (p=0.048).
CONCLUSION
Our study suggests that breast tumors containing EpCAM-/CD49f+ cells are more likely to undergo distant metastasis after initial surgery and are associated with a shorter DFS and OS.

Keyword

Breast neoplasms; Neoplasm metastasis; Neoplastic stem cells; Prognosis

MeSH Terms

Biomarkers
Breast Neoplasms*
Breast*
Disease-Free Survival
Fluorescent Antibody Technique
Humans
Immunohistochemistry
Multivariate Analysis
Neoplasm Metastasis
Neoplastic Stem Cells
Prognosis
Stem Cells

Figure

• 

  Figure 1 Dual-immunohistochemistry (IHC) and immunofluorescence (IF) staining on tissue microarray (TMA) slides. Dual-IHC and IF staining with primary antibodies of EpCAM and CD49f were performed on TMA tissue. (A) Patients only contain EpCAM-positive cancer cells (membrane); (B) Patient show only CD49f-positive cancer cells (cytoplasm); (C) Case express none or both (D) biomarkers. In IF staining (E-P), green signal shows the expression of CD49f, red signal shows EpCAM with DAPI staining for nuclei. (E-H) One case that tumor cell expression only EpCAM on the membrane (white arrows); (I-L) One case that tumor cell express only CD49f (red arrow); (M-P) One case with tumor cells express both (yellow arrow) (for both IHC and IF, magnification is actually ×400).

• 

  Figure 2 Prevalence of EpCAM-/CD49f+ with histopathologic characteristics. Analyzing of 150 breast cancer samples stratified to tumor size (A), node status (B), tumor grade (C; low=grade I and II, high=grade III), recurrence (D), distant metastasis (E), and estrogen receptor (ER) expression (F). Tumor containing EpCAM-/CD49f+ are associated with higher tumor grade (C; p=0.031) and distant metastasis (E; p=0.048). Chi-square exact and Fisher exact test were performed.

• 

  Figure 3 Univariate survival analysis of EpCAM-/CD49f+ with disease-free survival (DFS) and overall survival (OS). All the patients' tumor samples were stained with EpCAM and CD49f. (A, B) The present of EpCAM-/CD49f+ tumor cells have significant negative association with both DFS and OS (EpCAM-/CD49f+: pDFS=0.009; pOS=0.001).

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