Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin

Journal of Circulating Biomarkers
Joseph W PoTherese M Becker

Abstract

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 ...Continue Reading

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Citations

Apr 24, 2020·Cancer Medicine·Laan Luo, Yongqing He
Jun 7, 2020·International Journal of Molecular Sciences·Milad AshrafizadehGautam Sethi
Oct 12, 2019·NPJ Precision Oncology·Su Bin LimChwee Teck Lim
Dec 6, 2018·BioMed Research International·Wen-Chao WangXiao-Feng Zhang
Dec 2, 2020·Cancers·Peng LiuMichiel Stevens
Jul 20, 2019·Lung Cancer : Journal of the International Association for the Study of Lung Cancer·P N DingT L Roberts
Dec 29, 2020·Cancer Management and Research·Qiang ChiGuang Ma
Mar 4, 2021·Cancers·Amelia J RushtonR Charles Coombes
Sep 29, 2020·ACS Omega·Dhananjay SureshRaghuraman Kannan

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Methods Mentioned

BETA
biopsies
fluorescence-activated cell sorting
FACS
flow cytometry

Software Mentioned

Isoflux
CellSens Dimension
Flowing

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