The main focus of the study was to detect circulating tumor cells (CTCs) in ovarian cancer (OC) patients using a new methodological approach (MetaCell(TM)) which is based on size-dependent separation of CTCs and subsequent cytomorphological evaluation. Cytomorphological evaluation using vital fluorescence microscopy approach enables to use the captured cells for further RNA/DNA analysis. The cytomorphological analysis is then completed by gene expression analysis (GEA). GEA showed that relative expression of EPCAM is elevated in CTC-enriched fractions in comparison to the whole peripheral blood sample and that the expression grows with in vitro cultivation time. Comparison of the relative gene expression level in the group of peripheral blood samples and CTC-fraction samples confirmed a statistically significant difference for the following genes (p < 0.02): KRT7, WT1, EPCAM, MUC16, MUC1, KRT18 and KRT19. Thus, we suggest that the combination of the above listed genes could confirm CTCs presence in OC patients with higher specificity than when GEA tests are performed for one marker only. The GEA revealed two separate clusters identifying patients with or without CTCs.

Department of Laboratory Genetics University Hospital Kralovske Vinohrady Srobarova 50 10034 Prague Czech Republic

Department of Laboratory Genetics University Hospital Kralovske VinohradySrobarova 50 10034 Prague Czech Republic; Department of Pathology and Thoracic Surgery Masaryk's Hospital in Usti nad LabemKrajska zdravotni a s Socialni pece 3316 12A 40113 Usti nad Labem Czech Republic; Department of Histology and Embryology Wroclaw Medical Universitywybrzeże Ludwika Pasteura 1 50 367 Wrocław Poland; 3rd Department of Surgery University Hospital Motol and 1st Faculty of Medicine Charles UniversityV Uvalu 84 15006 Prague Czech Republic

Department of Pathology and Thoracic Surgery Masaryk's Hospital in Usti nad Labem Krajska zdravotni a s Socialni pece 3316 12A 40113 Usti nad Labem Czech Republic

Division of Surgical Oncology Gynaecological Oncology Chemotherapy and Department of Oncology Wroclaw Medical Universitywybrzeże Ludwika Pasteura 1 50 367 Wrocław Poland; Lower Silesian Cancer CenterWroclaw Plac Hirszfelda 12 53 413 Wrocław Poland

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