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Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin
Epithelial cell adhesion molecule (EpCAM)-targeted capture remains the most common isolation strategy for circulating tumor cells (CTCs). However, epithelial-to-mesenchymal transition (EMT) leads to decreased epithelial EpCAM expression affecting the optimal CTC capture. In this study, we tested a c...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6043919/ https://www.ncbi.nlm.nih.gov/pubmed/30013673 http://dx.doi.org/10.1177/1849454418782617 |
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author | Po, Joseph W Roohullah, Aflah Lynch, David DeFazio, Anna Harrison, Michelle Harnett, Paul R Kennedy, Catherine de Souza, Paul Becker, Therese M |
author_facet | Po, Joseph W Roohullah, Aflah Lynch, David DeFazio, Anna Harrison, Michelle Harnett, Paul R Kennedy, Catherine de Souza, Paul Becker, Therese M |
author_sort | Po, Joseph W |
collection | PubMed |
description | Epithelial cell adhesion molecule (EpCAM)-targeted capture remains the most common isolation strategy for circulating tumor cells (CTCs). However, epithelial-to-mesenchymal transition (EMT) leads to decreased epithelial EpCAM expression affecting the optimal CTC capture. In this study, we tested a cohort of ovarian cancer cell lines using flow cytometry to identify N-cadherin as the additional immunomagnetic cell surface target for ovarian cancer cell isolation. Combined immunomagnetic targeting of mesenchymal N-cadherin and epithelial EpCAM enriched CTCs from advanced ovarian cancer patient blood approximately three times more efficiently than targeting of EpCAM alone. We also show that more EMT-phenotype CTCs are captured by including N-cadherin targeting into CTC isolation protocols. However, after N-cadherin-based CTC isolation, in some blood samples of healthy individuals, we also observed the presence of cells expressing markers common to CTCs. Our data show that these “false positives” can be largely distinguished from CTCs as circulating endothelial cells (CECs) by vascular endothelial–cadherin co-staining. CEC counts are highly variable in patients and healthy controls. Our data demonstrate that a combination of EpCAM with N-cadherin-targeted isolation can improve CTC detection and widen the EMT-phenotype spectrum of captured CTCs. |
format | Online Article Text |
id | pubmed-6043919 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-60439192018-07-16 Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin Po, Joseph W Roohullah, Aflah Lynch, David DeFazio, Anna Harrison, Michelle Harnett, Paul R Kennedy, Catherine de Souza, Paul Becker, Therese M J Circ Biomark Research Article Epithelial cell adhesion molecule (EpCAM)-targeted capture remains the most common isolation strategy for circulating tumor cells (CTCs). However, epithelial-to-mesenchymal transition (EMT) leads to decreased epithelial EpCAM expression affecting the optimal CTC capture. In this study, we tested a cohort of ovarian cancer cell lines using flow cytometry to identify N-cadherin as the additional immunomagnetic cell surface target for ovarian cancer cell isolation. Combined immunomagnetic targeting of mesenchymal N-cadherin and epithelial EpCAM enriched CTCs from advanced ovarian cancer patient blood approximately three times more efficiently than targeting of EpCAM alone. We also show that more EMT-phenotype CTCs are captured by including N-cadherin targeting into CTC isolation protocols. However, after N-cadherin-based CTC isolation, in some blood samples of healthy individuals, we also observed the presence of cells expressing markers common to CTCs. Our data show that these “false positives” can be largely distinguished from CTCs as circulating endothelial cells (CECs) by vascular endothelial–cadherin co-staining. CEC counts are highly variable in patients and healthy controls. Our data demonstrate that a combination of EpCAM with N-cadherin-targeted isolation can improve CTC detection and widen the EMT-phenotype spectrum of captured CTCs. SAGE Publications 2018-06-24 /pmc/articles/PMC6043919/ /pubmed/30013673 http://dx.doi.org/10.1177/1849454418782617 Text en © The Author(s) 2018 http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Research Article Po, Joseph W Roohullah, Aflah Lynch, David DeFazio, Anna Harrison, Michelle Harnett, Paul R Kennedy, Catherine de Souza, Paul Becker, Therese M Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin |
title | Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of
epithelial EpCAM and mesenchymal N-cadherin |
title_full | Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of
epithelial EpCAM and mesenchymal N-cadherin |
title_fullStr | Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of
epithelial EpCAM and mesenchymal N-cadherin |
title_full_unstemmed | Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of
epithelial EpCAM and mesenchymal N-cadherin |
title_short | Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of
epithelial EpCAM and mesenchymal N-cadherin |
title_sort | improved ovarian cancer emt-ctc isolation by immunomagnetic targeting of
epithelial epcam and mesenchymal n-cadherin |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6043919/ https://www.ncbi.nlm.nih.gov/pubmed/30013673 http://dx.doi.org/10.1177/1849454418782617 |
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