Heterogeneous Manifestations of Epithelial–Mesenchymal Plasticity of Circulating Tumor Cells in Breast Cancer Patients

Abstract

To date, there is indisputable evidence of significant CTC heterogeneity in carcinomas, in particular breast cancer. The heterogeneity of CTCs is manifested in the key characteristics of tumor cells related to metastatic progression – stemness and epithelial–mesenchymal (EMT) plasticity. It is still not clear what markers can characterize the phenomenon of EMT plasticity in the range from epithelial to mesenchymal phenotypes. In this article we examine the manifestations of EMT plasticity in the CTCs in breast cancer. The prospective study included 39 patients with invasive carcinoma of no special type. CTC phenotypes were determined by flow cytometry before any type of treatment. EMT features of CTC were assessed using antibodies against CD45, CD326 (EpCam), CD325 (N-cadherin), CK7, Snail, and Vimentin. Circulating tumor cells in breast cancer are characterized by pronounced heterogeneity of EMT manifestations. The results of the study indicate that the majority of heterogeneous CTC phenotypes (22 out of 24 detectable) exhibit epithelial–mesenchymal plasticity. The variability of EMT manifestations does not prevent intravasation. Co-expression of EpCAM and CK7, regardless of the variant of co-expression of Snail, N-cadherin, and Vimentin, are associated with a low number of CTCs. Intrapersonal heterogeneity is manifested by the detection of several CTC phenotypes in each patient. Interpersonal heterogeneity is manifested by various combinations of CTC phenotypes in patients (from 1 to 17 phenotypes).

Circulating Tumor Cells from Enumeration to Analysis: Current Challenges and Future Opportunities

“…The CellSearch ® system technology (Menarini-Silicon Biosystems) Immunomagnetic cell selection system Antibody-labeled magnetic nanoparticles that target epithelial cell adhesion molecules of CTCs [29] AdnaTest Tumor cell enrichment based on magnetic beads that are coupled to a mixture of antibodies [30] MagSweeper…”

“…The isolation and capture of CTCs based on biological characteristics of those heterogeneous cells utilize different biological markers expressed in the cells ( Table 2 ) [ 18 , 19 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 45 , 56 , 57 , 58 , 59 , 60 , 61 , 62 ]. The first data describing the detection and characterization of carcinoma cells in the systemic circulation (bone marrow) based on cytokeratin and MUC-1 glycoprotein expression in epithelial cancer cells, but not in healthy cells, were published 25 years ago [ 63 ].…”

Generation of GCaMP6s-Expressing Zebrafish to Monitor Spatiotemporal Dynamics of Calcium Signaling Elicited by Heat Stress

“…To generate the transgenic construct pTol2-Cca.actb-GCaMP6s ( Figure 1 A), Tol2-elavl3-GCaMP6s obtained from Addgene (#59531, Watertown, MA, USA) and pT2/carp_actin_P-Abcc4-Flag-carp_actin_pA constructed in our previous study [ 61 ] were double digested using Hin d III and Eco R I from Fermentas (Burlington, NJ, Canada). The desired DNA fragments were recovered through electrophoresis and subsequent gel purification using a kit from Bioer (Hangzhou, China).…”

Heterogeneity of circulating epithelial cells in breast cancer at single-cell resolution: identifying tumor and hybrid cells

“…1a). CD45 ─ CECs were identified based on the expression of one or more epithelial markers: KRT 5,7,8,17,18,19, EpCAM, and CDH1 (E-cadherin) (Fig. 1b).…”

“…CTCs are highly heterogeneous, and only a few of them can survive and form metastases 5 . Many studies showed that breast CTCs possess specific stem and epithelialmesenchymal transition (EMT) properties [6][7][8][9][10] , while the molecular repertoire of CTCs varies from patient to patient 6 . Deciphering CTC heterogeneity is thought to be a vital issue for identifying metastasis-initiating cells and therapeutic targets for the prevention of metastasis.…”