INTRODUCTION: This 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. METHODS: Patients 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. RESULTS: CTC 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. DISCUSSIONS: Interestingly, 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-. CONCLUSIONS: Data 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.

Breast Unit Lower Silesian Cancer Center Wroclaw Plac Hirszfelda 12 53 413 Wrocław Poland

Department of Histology and Embryology Wroclaw Medical University Wybrzeże Ludwika Pasteura 1 50 367 Wrocław Poland

Department of Laboratory Genetics University Hospital Kralovske Vinohrady Srobarova 50 100 34 Prague Czech Republic

Department of Oncology 1st Faculty of Medicine Charles University and General University Hospital Prague Prague Czech Republic

Department of Surgery University Hospital Motol 5 Uvalu 84 150 06 Prague Czech Republic

Department of Thoracic Surgery Masaryk´s Hospital Krajska zdravotni a s Socialni pece 3316 12A 40113 Usti Nad Labem Czech Republic

Division of Surgical Oncology and Department of Oncology Wroclaw Medical University Wybrzeże Ludwika Pasteura 1 50 367 Wrocław Poland

Faculty of Medicine Charles University 5 Uvalu 84 150 06 Prague Czech Republic

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