J Breast Cancer.  2012 Mar;15(1):15-23. 10.4048/jbc.2012.15.1.15.

Aberrant Expression of Cancer Stem Cells Marker Prominin-1 in Low-Grade Tubulobular Breast Carcinoma: A Correlative Study between qRT-PCR, Flow-Cytometric and Immunohistochemistry Analysis

• Di Bonito
• Collina F
• Cantile M
• Camerlingo R
• Cerrone M
• Marra L
• Liguori G
• Pirozzi G
• Botti G

• KMID: 2286469
• DOI: http://doi.org/10.4048/jbc.2012.15.1.15

Abstract

PURPOSE
Prominin1/CD133 has become the ideal marker for cancer stem cells (CSCs) detection in human tumors. In this study we examined the expression of this marker in several breast cancer specimens to associate CSCs percentage with risk factor for this neoplasia.
METHODS
We examined specimens from 12 patients using CD133 and CD44 antibodies for CSCs immunohistochemistry detection and for flow cytometry analysis. For each patient, we also performed the immunohistochemical staining to evaluate the expression of estrogen receptor, progesterone receptor, c-erbB-2, Ki67, and E-cadherin markers. A Taqman probe for CD133 was used for mRNA quantification by real-time polymerase chain reaction.
RESULTS
Prominin-1 expression was heterogeneous in different carcinomas but was strikingly hyperexpressed in a tubulolobular variant of breast cancer. The results were confirmed by all three methods.
CONCLUSION
Our data, although produced on a limited number of samples, showed an particularly high expression of stem cell marker CD133 in a breast cancer variant, generally with a good prognosis. Since CSCs detection by CD133 has been described as an important prognostic factor for several human cancers, we suggest the importance of detecting stem cell compartiments in all histotypes of breast carcinomas.

Keyword

CD133; Neoplastic stem cells; Tubulobular breast cancer

MeSH Terms

Antibodies
Antigens, CD
Breast
Breast Neoplasms
Cadherins
Estrogens
Flow Cytometry
Glycoproteins
Humans
Immunohistochemistry
Neoplastic Stem Cells
Peptides
Prognosis
Receptors, Progesterone
Risk Factors
RNA, Messenger
Stem Cells
Antibodies
Antigens, CD
Cadherins
Estrogens
Glycoproteins
Peptides
RNA, Messenger
Receptors, Progesterone

Figure

• 

  Figure 1 Tubulolobular breast immunophenotype. (A) H&E morphology (×20). (B) immunopositivity for progesterone receptor (×60). (C) immunopositivity for estrogen receptor (×60). (D) immunonegativity for HER2/neu (×60). (E) immunopositivity for Ki67 (×60). (F) immunopositivity for E-Cadherin in ductal cells and immunonegativity in lobular cells (×60).

• 

  Figure 2A CD133 and CD44 immunostaining: on the left in the first (CD133) and second (CD44) columns immunohistochemistry detection in sample 1, 2, 3, 4, 5, 6 (×60); on the right in third (CD133) and fourth (CD44) columns histogram plot representation of flow cytometry detection in sample 1, 2, 3, 4, 5, 6.

• 

  Figure 2B CD133 and CD44 immunostaining: on the left in the first (CD133) and second (CD44) columns immunohistochemistry detection in samples 7, 9, 10, 11, 12 (×60); on the right in third (CD133) and fourth (CD44) columns histogram plot representation of flow cytometry detection in samples 7, 9, 10, 11, 12.

• 

  Figure 3 CD133 and CD44 immunostaining in tubulolobular breast variant. (A) Immunopositivity for CD133 (×40). (B) immunopositivity for CD133 (×60). (C) immunopositivity for CD133 in tubular and lobular components (×60). (D) immunopositivity for CD44 (×60). (E) immunonegativity for CD133 in normal ductal cells (×40). (F) immunopositivity for CD44 in normal ductal cells (×40).

• 

  Figure 4 CD133 and CD44 flow cytometry detection in tubulolobular breast variant. (A, C) Control. (B, D) CD133 expression in sample 8. (E, F) CD44 expression in sample 8.

• 

  Figure 5 Prominin-1/CD133 real-time expression in breast samples. All reactions were performed in triplicate and data are expressed as mean of relative amount of mRNAs levels.

• 

  Figure 6 CD133 immunostaining in archive tubulolobular breast variant. (13, 17, 20) immunopositivity for CD133 (×40). (14, 15, 16, 18, 19) immunonegativity for CD133 (×40).

• 

  Figure 7 Prominin-1/CD133 real-time expression in archive tubulolobular breast samples. All reactions were performed in triplicate and data are expressed as mean of relative amount of mRNAs levels.

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