Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells

AbstractApplication of tumor cell surface adhesion molecule EpCAM‐dependent antibody capture, and intracellular cytokeratins (CKs)‐dependent immunostaining strategies to detect disseminated or circulating tumor cells (DTCs or CTCs), is limited by highly heterogeneous and dynamic expression or absenc...

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Main Author: Peter Ping Lin
Format: Article
Language:English
Published: Wiley 2015-12-01
Series:Clinical and Translational Medicine
Subjects:
Online Access:https://doi.org/10.1186/s40169-015-0081-2
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author Peter Ping Lin
author_facet Peter Ping Lin
author_sort Peter Ping Lin
collection DOAJ
description AbstractApplication of tumor cell surface adhesion molecule EpCAM‐dependent antibody capture, and intracellular cytokeratins (CKs)‐dependent immunostaining strategies to detect disseminated or circulating tumor cells (DTCs or CTCs), is limited by highly heterogeneous and dynamic expression or absence of EpCAM and/or CKs in CTCs and DTCs, particularly in their capturing and identifying CTCs/DTCs shed from diverse types of solid tumor, thus being biased and restricted to the only both EpCAM and CK positive cancer cells. Moreover, heterogeneity of chromosome and tumor biomarker of CTCs/DTCs cannot be co‐examined by conventional CK/EpCAM‐dependent techniques. Accordingly, a novel integrated cellular and molecular approach of EpCAM‐independent subtraction enrichment (SE) and immunostaining‐FISH (iFISH®) has recently been successfully developed. SE‐iFISH® is able to effectively enrich, comprehensively identify and characterize both large and small size non‐hematopoietic heteroploid CTCs, DTCs and circulating tumor microemboli in various biofluid specimens of either cancer patients or patient‐derived‐xenograft mice. Obtained tumor cells, free of anti‐EpCAM perturbing and hypotonic damage, are eligible for primary tumor cell culture as well as a series of downstream analyses. Highly heterogeneous CTCs and DTCs could be classified into subtypes by in situ phenotyping protein expression of various tumor biomarkers and karyotyping of chromosome aneuploidy performed by iFISH®. Each CTC subtype may correlate with distinct clinical significance in terms of tumor metastasis, relapse, therapeutic drug sensitivity or resistance, respectively.
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spelling doaj.art-c432e74e82b5499a920c504257521a1d2022-12-21T22:52:05ZengWileyClinical and Translational Medicine2001-13262015-12-0141n/an/a10.1186/s40169-015-0081-2Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cellsPeter Ping Lin0Cytelligen92121San DiegoCAUSAAbstractApplication of tumor cell surface adhesion molecule EpCAM‐dependent antibody capture, and intracellular cytokeratins (CKs)‐dependent immunostaining strategies to detect disseminated or circulating tumor cells (DTCs or CTCs), is limited by highly heterogeneous and dynamic expression or absence of EpCAM and/or CKs in CTCs and DTCs, particularly in their capturing and identifying CTCs/DTCs shed from diverse types of solid tumor, thus being biased and restricted to the only both EpCAM and CK positive cancer cells. Moreover, heterogeneity of chromosome and tumor biomarker of CTCs/DTCs cannot be co‐examined by conventional CK/EpCAM‐dependent techniques. Accordingly, a novel integrated cellular and molecular approach of EpCAM‐independent subtraction enrichment (SE) and immunostaining‐FISH (iFISH®) has recently been successfully developed. SE‐iFISH® is able to effectively enrich, comprehensively identify and characterize both large and small size non‐hematopoietic heteroploid CTCs, DTCs and circulating tumor microemboli in various biofluid specimens of either cancer patients or patient‐derived‐xenograft mice. Obtained tumor cells, free of anti‐EpCAM perturbing and hypotonic damage, are eligible for primary tumor cell culture as well as a series of downstream analyses. Highly heterogeneous CTCs and DTCs could be classified into subtypes by in situ phenotyping protein expression of various tumor biomarkers and karyotyping of chromosome aneuploidy performed by iFISH®. Each CTC subtype may correlate with distinct clinical significance in terms of tumor metastasis, relapse, therapeutic drug sensitivity or resistance, respectively.https://doi.org/10.1186/s40169-015-0081-2CTC and DTC subtypeHeteroploid chromosomeTumor biomarkerMetastasisLiquid biopsy
spellingShingle Peter Ping Lin
Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
Clinical and Translational Medicine
CTC and DTC subtype
Heteroploid chromosome
Tumor biomarker
Metastasis
Liquid biopsy
title Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
title_full Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
title_fullStr Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
title_full_unstemmed Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
title_short Integrated EpCAM‐independent subtraction enrichment and iFISH strategies to detect and classify disseminated and circulating tumors cells
title_sort integrated epcam independent subtraction enrichment and ifish strategies to detect and classify disseminated and circulating tumors cells
topic CTC and DTC subtype
Heteroploid chromosome
Tumor biomarker
Metastasis
Liquid biopsy
url https://doi.org/10.1186/s40169-015-0081-2
work_keys_str_mv AT peterpinglin integratedepcamindependentsubtractionenrichmentandifishstrategiestodetectandclassifydisseminatedandcirculatingtumorscells