Phenotypic and Genetic Characterization of Circulating Tumor Cells by Combining Immunomagnetic Selection and FICTION Techniques

Author:

Campos María1,Prior Celia2,Warleta Fernando1,Zudaire Isabel3,Ruíz-Mora Jesús1,Catena Raúl24,Calvo Alfonso24,José J. Gaforio1

Affiliation:

1. Immunology Division, Department of Health Sciences, Faculty of Experimental Sciences, University of Jaén, Jaén, Spain

2. Division of Oncology, Center for Applied Medical Research (CIMA), Pamplona, Spain

3. Department of Genetics, University of Navarra, Pamplona, Spain

4. Department of Histology and Pathology, University of Navarra, Pamplona, Spain

Abstract

The presence of circulating tumor cells (CTCs) in breast cancer patients has been proven to have clinical relevance. Cytogenetic characterization of these cells could have crucial relevance for targeted cancer therapies. We developed a method that combines an immunomagnetic selection of CTCs from peripheral blood with the fluorescence immunophenotyping and interphase cytogenetics as a tool for investigation of neoplasm (FICTION) technique. Briefly, peripheral blood (10 ml) from healthy donors was spiked with a predetermined number of human breast cancer cells. Nucleated cells were separated by double density gradient centrifugation of blood samples. Tumor cells (TCs) were immunomagnetically isolated with an anti-cytokeratin antibody and placed onto slides for FICTION analysis. For immunophenotyping and genetic characterization of TCs, a mixture of primary monoclonal anti-pancytokeratin antibodies was used, followed by fluorescent secondary antibodies, and finally hybridized with a TOP2A/HER-2/CEP17 multicolor probe. Our results show that TCs can be efficiently isolated from peripheral blood and characterized by FICTION. Because genetic amplification of TOP2A and ErbB2 (HER-2) in breast cancer correlates with response to anthracyclines and herceptin therapies, respectively, this novel methodology could be useful for a better classification of patients according to the genetic alterations of CTCs and for the application of targeted therapies.

Publisher

SAGE Publications

Subject

Histology,Anatomy

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