Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels

A. Bellahcène, N. Antoine, N. Clausse, E. Tagliabue, L. W. Fisher, J. M. Kerr, P. Jarès, V. Castronovo

Research output: Contribution to journalArticle

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Abstract

The recent demonstration that bone sialoprotein (BSP) can be detected in human breast cancer tissue by immunoperoxidase suggests that this phosphoprotein is ectopically expressed by malignant mammary epithelial cells. Its detection in human breast cancer cells raises questions about its potential role(s) during breast cancer progression. Because BSP is secreted and is present in the serum, the positivity of breast cancer cells for BSP could have been the result of an uptake of the circulating phosphoprotein by the cells rather than of an intrinsic expression. We examined the expression of BSP at both the protein and mRNA levels in nine human breast cancer samples as well as in three human breast cancer cell lines (MCF-7, T47-D, and MDA-MB-231) using immunohistochemistry, flow cytometric analysis, immunoblot, and reverse-transcriptase PCR. BSP was detected at both protein and mRNA levels in human breast cancer tissue and in the three human breast cancer cell lines. Using a specific polyclonal anti-BSP antibody, we showed by both fluorescence-activated cell sorter analysis and immunohistochemistry experiments that all of the human breast cancer cell lines studied express BSP. This was localized at the cell surface and in the cytosol of the estrogen receptor-positive MCF-7 and T47-D cell lines, whereas it was detected only in the cytosol of the estrogen receptor-negative MDA-MB-231 cells. Using the same polyclonal anti-BSP antibody, we were able to identify an approximately 97-kd band on total protein extracts from the three cell lines by immunoblotting. Reverse-transcriptase PCR reactions using specific oligonucleotides performed on total RNA of nine human breast cancer biopsy samples and the three cell lines demonstrated the presence of BSP mRNA in all of the samples examined. This study is the first demonstration that human malignant breast epithelial cell lines express BSP at the protein and mRNA levels. Our study identified MCF-7, T47-D, and MDA-MB-231 cells as useful models for the examination of the molecular mechanisms involved in the ectopic expression of BSP in breast malignant lesions.

Original languageEnglish
Pages (from-to)203-210
Number of pages8
JournalLaboratory Investigation
Volume75
Issue number2
Publication statusPublished - Aug 1996

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Integrin-Binding Sialoprotein
RNA
Breast Neoplasms
Cell Line
Proteins
Messenger RNA
Breast
Phosphoproteins
Reverse Transcriptase Polymerase Chain Reaction
Estrogen Receptors
Cytosol
Epithelial Cells
Immunohistochemistry
Bone Neoplasms
Molecular Models
Somatostatin-Secreting Cells
Antibodies
Immunoblotting
Oligonucleotides

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Bellahcène, A., Antoine, N., Clausse, N., Tagliabue, E., Fisher, L. W., Kerr, J. M., ... Castronovo, V. (1996). Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels. Laboratory Investigation, 75(2), 203-210.

Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels. / Bellahcène, A.; Antoine, N.; Clausse, N.; Tagliabue, E.; Fisher, L. W.; Kerr, J. M.; Jarès, P.; Castronovo, V.

In: Laboratory Investigation, Vol. 75, No. 2, 08.1996, p. 203-210.

Research output: Contribution to journalArticle

Bellahcène, A, Antoine, N, Clausse, N, Tagliabue, E, Fisher, LW, Kerr, JM, Jarès, P & Castronovo, V 1996, 'Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels', Laboratory Investigation, vol. 75, no. 2, pp. 203-210.
Bellahcène, A. ; Antoine, N. ; Clausse, N. ; Tagliabue, E. ; Fisher, L. W. ; Kerr, J. M. ; Jarès, P. ; Castronovo, V. / Detection of bone sialoprotein in human breast cancer tissue and cell lines at both protein and messenger ribonucleic acid levels. In: Laboratory Investigation. 1996 ; Vol. 75, No. 2. pp. 203-210.
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