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.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-015-9866-9) contains supplementary material, which is available to authorized users.
Highlights
Metastatic dissemination is an important prognostic factor for patients with gastro-intestinal cancer
Follow-up studies in gastric cancer (GC) patients suggest that CTCpositive cases with an increased burden of Circulating Tumor Cells (CTCs) were associated with a poorer prognosis than CTC-negative patients, and the situation was similar for disseminated tumor cells (DTCs) (Wang et al 2009)
We report successful CTC isolation in 59 % of GC patients (n = 13/22)
Summary
Metastatic dissemination is an important prognostic factor for patients with gastro-intestinal cancer. Follow-up studies in GC patients suggest that CTCpositive cases with an increased burden of CTCs were associated with a poorer prognosis than CTC-negative patients, and the situation was similar for DTCs (Wang et al 2009). Both localized and metastatic GC can shed a detectable concentration of CTCs into the blood. The presence of CTCs in the circulation indicates a high risk of tumor recurrence as well as unfavourable clinical outcomes, even for early GC (Zhang and Ge 2013). In this study we demonstrate its use for enrichment, separation and cultivation of CTCs
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