Simultaneous phenotypic and genetic characterization of single circulating tumor cells from colon cancer patients
María Campos1, Rafael Luque2, Juan Jiménez3, Rafael Martínez4, Fernando Warleta1, Cristina Sánchez-Quesada1, Miguel Delgado-Rodríguez5,6, Alfonso Calvo7 and José J. Gaforio1
1Immunology Division, Department of Health Sciences, Faculty of Experimental Sciences, University of Jaén, Campus Las Lagunillas Jaén, 2Departments of Pathology, 3Surgery, 4Gastroenterology, University Hospital of Jaén, Jaén, 5Division of Preventive Medicine and Public Health, Department of Health Sciences, Faculty of Experimental Sciences, University of Jaén, Campus Las Lagunillas, Jaén, 6Biomedical Network Research Center for Epidemiology and Public Health (CIBERESP), Ministry of Science and Innovation and 7Division of Oncology, Center for Applied Medical Research (CIMA) and Department of Histology and Pathology, University of Navarra, Pamplona, Spain.
Offprint requests to: José J. Gaforio, Immunology Division Department of Health Sciences, Faculty of Experimental Sciences, University of Jaén, Campus Las Lagunillas s/n, 23071 Jaén, Spain. e-mail: email@example.com
Summary. Since circulating tumor cells (CTCs) have metastatic potential, their genetic and phenotypic characteristics could provide crucial information to establish the most effective therapy. We assessed the clinical utility of a methodology that allows the simultaneous analysis of CTC phenotype and genotype in colon cancer patients and, in addition, whether this methodology could provide complementary information to that obtained by the primary tumor biopsy.
Thirty-three non-metastatic (stages 0-III) colon cancer patients and 9 healthy donor samples were evaluated. All peripheral blood samples (10 ml) were analyzed by cytokeratin immunomagnetic enrichment. Eight samples were analyzed by immunocytochemistry and 25 samples were analyzed by FICTION technique for simultaneous cytokeratin expression and chromosome 17 and ERBB2 gene status. A further study was carried out in one patient who showed CTC heterogeneity in chromosomal abnormalities. We analyzed HER2 protein expression on CTCs and FISH and HER2 protein expression in primary tumor of this patient.
Our results show that 9.09% of patients had cytokeratin-positive CTCs (CK+/CTCs in peripheral blood). One of the patients showed heterogeneity in chromosomal 17 abnormalities and two different CK expression patterns on CTCs: one CK+/CTCs and one CK-/CTCs. Furthermore, 63.33% of these CTCs overexpressed HER2 protein while the primary tumor of this patient was diploid and did not express HER2 protein.
We describe a methodology that allows the simultaneous genetic and phenotypic analysis of CTCs in colon cancer patients, which may provide essential information to select patients who might benefit from specific therapy. Histol Histopathol 28, 1439-1450 (2013)
Key words: CK+/CTCs, Colon cancer, ERBB2, FICTION, HER2