IntroductionThis study analyzes peripheral blood samples from breast cancer (BC) patients. CTCs from peripheral blood were enriched by size-based separation and were then cultivated in vitro. The primary aim of this study was to demonstrate the antigen independent CTC separation method with high CTC recovery. Subsequently, CTCs enriched several times during the treatment were characterized molecularly.MethodsPatients with different stages of BC (N = 167) were included into the study. All patients were candidates for surgery, surgical diagnostics, or were undergoing chemotherapy. In parallel, 20 patients were monitored regularly and in addition to CTC presence, also CTC character was examined by qPCR, with special focus on HER2 and ESR status.ResultsCTC positivity in the cohort was 76%. There was no significant difference between the tested groups, but the highest CTC occurrence was identified in the group undergoing surgery and similarly in the group before the start of neoadjuvant treatment. On the other hand, the lowest CTC frequencies were observed in the menopausal patient group (56%), ESR+ patient group (60%), and DCIS group (44.4%). It is worth noting that after completion of neoadjuvant therapy (NACT) CTCs were present in 77.7% of cases. On the other hand, patients under hormonal treatment were CTC positive only in 52% of cases.DiscussionsInterestingly, HER2 and ESR status of CTCs differs from the status of primary tumor. In 50% of patients HER2 status on CTCs changed not only from HER2+ to HER2−, but also from HER2– to HER2+ (33%). ESR status in CTCs changed only in one direction from ESR+ to ESR−.ConclusionsData obtained from the present study suggest that BC is a heterogeneous disease but CTCs may be detected independently of the disease characteristics in 76% of patients at any time point during the course of the disease. This relatively high CTC occurrence in BC should be considered when planning the long-term patient monitoring.
Introduction. Liquid biopsies are noninvasive tests using blood or body fluids to detect circulating tumor cells (CTCs) or the products of tumor cells, such as fragments of nucleic acids or proteins that are shed into biological fluids from primary tumor or its metastates. The analysis of published clinical studies provides coherent evidence that the presence of CTCs detected in peripheral blood is a strong prognostic factor in patients with colorectal carcinoma (CRC). The aim of the study was to implement size-based separation protocol of CTCs in CRC patients. Material and methods. Patients diagnosed with different stages of CRC (n = 98) were included in the study. All patients have been diagnosed for colorectal adenocarcinoma by pathology examination, 45 patients with colon carcinoma and 53 with rectosigmoid cancer. A size-based separation method (MetaCell ® ) for viable CTC enrichment from peripheral blood was used to assess the presence of CTCs by cytomorphological evaluation using vital fluorescence microscopy. Results. Cytomorphological analysis revealed that 81 (83%) tested samples were CTC-positive and 17 (17%) were CTC-negative. We report a successful isolation of CTCs with proliferation potential in patients with CRC. The CTCs were cultured in vitro for further downstream applications. Some of the isolated CTCs were able to grow in vitro for 6 months as a standard cell culture. Conclusions. We established a reliable, inexpensive and relatively fast protocol for CTCs enrichment in CRC patients by means of vital fluorescence staining which enables their further analysis in vitro.
Abstract:Circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) are responsible for the development of metastatic disease, and may also hold the key to determining tailored therapies of advanced cancer disease. Our review summarizes the prognostic significance of the detection of CTCs and DTCs in various gastrointestinal cancers with an overview of their possible use as prognostic biomarkers. This could be used in the future as a starting point for new clinical trials focusing on the predictive potential of circulating and disseminated tumor cells. (Folia Histochemica et Cytobiologica 2013, Vol. 51, No. 4, 265-277) Key words: circulating tumor cells; gastrointestinal cancer; esophageal cancer; colorectal cancer; gastric cancer; plastin3; prognosis Abbreviations AFP -alpha fetoprotein; BM -bone marrow; CD -cluster of differentiation; CEA -carcinoembryonic antigen; CHT -chemotherapy; CI -confidence interval; CTC -circulating tumor cell; CRCcolorectal carcinoma; CVB -central venous blood; CK -cytokeratin; DAPI -4,6-diamidino-2-phenylindole; DFS -disease free survival; DTC -disseminated tumor cell; EpCAM -epithelial cell adhesion molecule; FISH -fluorescent in situ hybridization; 5-FU -5-fluorouracil; HCC -hepatocellular carcinoma; HR -hazard ratio; ISET -isolation by size of epithelial tumor; ITC -isolated tumor cells; MACS -magnetic activated cell sorting; MFS -metastasis free survival; MSP -methylation specific polymerase chain reaction; MVB -mesenteric venous blood; NA -not available; OS -overall survival; PB -peripheral blood; PFS -progression-free survival; qPCR -quantitative real-time polymerase chain reaction; RFA -radiofrequency ablation; RT -radiotherapy; RT-PCR -reverse transcription polymerase chain reaction; TGFb1 -transforming growth factor b1; TRC method -transcription reverse-transcription concerted method
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