title: Detection and cultivation of circulating tumor cells in gastric cancer.
abstract: Circulating tumor cells (CTCs) are important targets for treatment and critical surrogate markers when evaluating cancer prognosis and therapeutic response. A sensitive methodology for detecting CTCs in gastric cancer (GC) patients is needed. In this study we demonstrate a device for enrichment and cultivation of CTCs. In total, 22 patients with GC, all candidates for surgery, were enrolled in the study. Peripheral blood samples were collected before surgery, and patients were re-evaluated within operation and divided into two groups: resectable and non-resectable GC. A new size-based separation test for enrichment and cultivation of CTCs was used (MetaCell(®)). In addition to cytomorphological analysis, gene expression of tumor associated genes (Cytokeratin-18, Cytokeratin-19, Cytokeratin-20, Cytokeratin-7, EPCAM, MUC1, HER2, EGFR) and of leukocyte markers (e.g. CD45, CD68) was tested in enriched CTC fractions. CTCs were detected in 59 % of the patients studied (n = 13/22). CTCs were detected in seven patients of the resection group (7/10, 70 %) and six of the non-resectable group (6/12, 50 %). Enrichment of the viable CTCs allowed subsequent successful cultivation in vitro. The cytomorphological characterization of the CTCs was a prerequisite of random gene expression testing in CTC-positive samples. In CTC-positive samples gene expression of cytokeratin 18 and 19 was elevated in comparison to the whole blood gene expression analysis. CTCs were found to be present in both resectable and non-resectable gastric cancer patients. The size-based separation platform for CTCs may be used for in vitro cultivation, as well as in subsequent molecular analysis if desired. The sensitivity of CTC-detection could be enhanced by the combination of cytomorphological and molecular analysis.
authors: Department of Laboratory Genetics, University Hospital, Kralovske Vinohrady, Srobarova 50, 100 34, Prague, Czech Republic.
authors: Katarina
authors: K
authors: Kolostova
authors: Division of Oncological Surgery, Department of Oncology, Wroclaw Medical University, Plac Hirszfelda 12, 53-413, Wrocław, Poland.|Lower Silesian Oncology Centre, Plac Hirszfelda 12, 53-413, Wrocław, Poland.
authors: Rafal
authors: R
authors: Matkowski
authors: Department of Surgery, 3rd Faculty of Medicine, Charles University Prague and University Hospital Kralovske Vinohrady, Srobarova 50, Prague, Czech Republic.
authors: Robert
authors: R
authors: Gürlich
authors: Department of Gastrointestinal and General Surgery, Wroclaw Medical University, ul. M. Curie-Skłodowskiej 66, 50-369, Wrocław, Poland.
authors: Krzysztof
authors: K
authors: Grabowski
authors: Division of Oncological Surgery, Department of Oncology, Wroclaw Medical University, Plac Hirszfelda 12, 53-413, Wrocław, Poland.
authors: Katarzyna
authors: K
authors: Soter
authors: 3rd Department of Surgery, First Faculty of Medicine, Charles University in Prague and University Hospital Motol, Prague, Czech Republic.
authors: Robert
authors: R
authors: Lischke
authors: 3rd Department of Surgery, First Faculty of Medicine, Charles University in Prague and University Hospital Motol, Prague, Czech Republic.
authors: Jan
authors: J
authors: Schützner
authors: Department of Laboratory Genetics, University Hospital, Kralovske Vinohrady, Srobarova 50, 100 34, Prague, Czech Republic. vbobek@centrum.cz.|3rd Department of Surgery, First Faculty of Medicine, Charles University in Prague and University Hospital Motol, Prague, Czech Republic. vbobek@centrum.cz.|Department of Histology and Embryology, Wroclaw Medical University, Chalubinskiego 6a, 50-368, Wroclaw, Poland. vbobek@centrum.cz.
authors: Vladimir
authors: V
authors: Bobek
keywords: CTC; Circulating tumor cells; Cultivation; Gastric cancer; Metacell