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Breast Cancer Research
Published in: Breast Cancer Research 3/2009

Open Access 01-06-2009 | Research article

Full-length cytokeratin-19 is released by human tumor cells: a potential role in metastatic progression of breast cancer

Authors: Catherine Alix-Panabières, Jean-Pierre Vendrell, Monique Slijper, Olivier Pellé, Eric Barbotte, Grégoire Mercier, William Jacot, Michel Fabbro, Klaus Pantel

Published in: Breast Cancer Research | Issue 3/2009

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Abstract

Introduction

We evaluated whether CK19, one of the main cytoskeleton proteins of epithelial cells, is released as full-length protein from viable tumor cells and whether this property is relevant for metastatic progression in breast cancer patients.

Methods

EPISPOT (EPithelial ImmunoSPOT) assays were performed to analyze the release of full-length CK19 by carcinoma cells of various origins, and the sequence of CK19 was analyzed with mass spectrometry. Additional functional experiments with cycloheximide, Brefeldin A, or vincristine were done to analyze the biology of the CK19-release. CK19-EPISPOT was used to detect disseminated tumor cells in bone marrow (BM) of 45 breast cancer patients who were then followed up over a median of 6 years.

Results

CK19 was expressed and released by colorectal (HT-29, HCT116, Caco-2) and breast (MCF-7, SKBR3, and MDA-MB-231) cancer cell lines. The CK19-EPISPOT was more sensitive than the CK19-ELISA. Dual fluorescent EPISPOT with antibodies against different CK19 epitopes showed the release of the full-length CK19, which was confirmed by mass spectrometry. Functional experiments indicated that CK19 release was an active process and not simply the consequence of cell death. CK19-releasing cells (RCs) were detectable in BM of 44% to 70% of breast cancer patients. This incidence and the number of CK19-RCs were correlated to the presence of overt metastases, and patients with CK19-RCs had a reduced survival as compared with patients without these cells (P = 0.025, log-rank test; P = 0.0019, hazard ratio, 4.7; multivariate analysis).

Conclusions

Full-length CK19 is released by viable epithelial tumor cells, and CK19-RCs might constitute a biologically active subset of breast cancer cells with high metastatic properties.
Appendix
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Metadata
Title
Full-length cytokeratin-19 is released by human tumor cells: a potential role in metastatic progression of breast cancer
Authors
Catherine Alix-Panabières
Jean-Pierre Vendrell
Monique Slijper
Olivier Pellé
Eric Barbotte
Grégoire Mercier
William Jacot
Michel Fabbro
Klaus Pantel
Publication date
01-06-2009
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 3/2009
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr2326

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