Feasibility of a novel one-stop ISET device to capture CTCs and its clinical application

Abstract

IntroductionCirculating tumor cells (CTCs) play a crucial role in cancer metastasis. In this study, we introduced a novel isolation method by size of epithelial tumor cells (ISET) device with automatic isolation and staining procedure, named one-stop ISET (osISET) and validated its feasibility to capture CTCs from cancer patients. Moreover, we aim to investigate the correlation between clinicopathologic features and CTCs in colorectal cancer (CRC) in order to explore its clinical application.ResultsThe capture efficiency ranged from 80.3% to 88% with tumor cells spiked into medium while 67% to 78.3% with tumor cells spiked into healthy donors’ blood. In detection blood samples of 72 CRC patients, CTCs and clusters of circulating tumor cells (CTC-clusters) were detected with a positive rate of 52.8% (38/72) and 18.1% (13/72) respectively. Moreover, CTC positive rate was associated with factors of lymphatic or venous invasion, tumor depth, lymph node metastasis and TNM stage in CRC patients (p < 0.01). Lymphocyte count and neutrophil to lymphocyte ratio (NLR) were significantly different between CTC positive and negative groups (p < 0.01).Materials and MethodsThe capture efficiency of the device was tested by spiking cancer cells (MCF-7, A549, SW480, Hela) into medium or blood samples of healthy donors. Blood samples of 72 CRC patients were detected by osISET device. The clinicopathologic characteristics of 72 CRC patients were collected and the association with CTC positive rate or CTC count were analyzed.ConclusionsOur osISET device was feasible to capture and identify CTCs and CTC-clusters from cancer patients. In addition, our device holds a potential for application in cancer management.

Combined Features Based on Preoperative Controlling Nutritional Status Score and Circulating Tumour Cell Status Predict Prognosis for Colorectal Cancer Patients Treated with Curative Resection

“…It has been estimated that approximately 1×106 CTCs per gram of tumour tissue are released into the circulation daily [14], but most of them are recognized and cleared by immune cells in the blood microenvironment; ultimately, less than 0.1% CTCs can reach distant organs to form metastasis lesion [15]. The above perspective has also been confirmed in our previous studies, which showed that the number of CTCs was negatively correlated with lymphocyte count in peripheral blood [16,17]. Of note, we also found that CTC count had a negative correlation with decreased prognostic nutrition index (PNI) [18], which is an important indicator reflecting the perioperative nutritional conditions of patients with malignant gastrointestinal tract tumours [19].…”

“…CTC isolation was performed using the CTCBIOPSY® device (Wuhan YZY Medical Science and Technology Co., Ltd., Wuhan, China), as described in our previous studies [11,16]. The specific implementation was completed by the professional staff of Wuhan YZY Medical Science and Technology Company according to the manufacturer's instruction.…”

“…In brief, 5 ml peripheral blood samples from each patient were collected in EDTA-containing tubes (BD, USA) and diluted up to 15 ml with 0.9% physiological saline containing 0.2% paraformaldehyde; then, the samples were transferred to tubes with an 8 μm diameter aperture membrane. After being filtered by positive pressure from 12 to 20 mmHg, the candidate CTCs adhered to the membrane and were identified by three-colour immunofluorescence staining, as described in our previous study [16]. Briefly, membranes with CTC were transferred to glass slides and were fixed with 4% PFA for 5 minutes.…”

Significant diagnostic value of circulating tumour cells in colorectal cancer

“…This strategy achieved a diagnostic sensitivity of 83.05% and a specificity of 100%, when using a cut-off value of 2 CTCs/3.2 ml of peripheral blood. The detection rate of CTCs with this method was higher than that of the CellSearch ® system, which has been reported to be 15.33% (10), higher than that of the ISET ® method, which has been reported to be 35% (20), and higher than that of the approach using RT-qPCR and ECT2, which has been reported to be 59% (21). Furthermore, the Cyttel method requires less peripheral blood (3.2 ml) than the CellSearch ® system (7 ml).…”

“…The methods for CTC detection have been recently improved. Chen et al (20) introduced a novel isolation method, known as the ISET ® system, involving automatic isolation and staining procedures to capture CTCs, and revealed that CTCs were detected in 38/72 (52.8%) patients with CRC. In another study based on nested RT-qPCR, epithelial cell transforming 2 (ECT2) was screened as a candidate marker gene for quantifying CTCs, with a detection rate of ECT2 of ~60% in patients with different stages of CRC (21).…”