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ORIGINAL RESEARCH
Characteristics of the metastasis-associated circulating cells: features of side scatter parameters
1 Cancer Research Institute, Tomsk National Research Medical Center of the Russian Academy of Sciences, Tomsk, Russia
2 Siberian State Medical University, Tomsk, Russia
3 Saint Petersburg State Pediatric Medical University, Saint Petersburg, Russia
Correspondence should be addressed: Angelina V. Buzenkova
Kooperativny per., 5, Tomsk, 634009, Russia; ur.liam@va_avoknezub
Funding: the study was supported by the RSF grant No. № 23-15-00135.
Author contribution: Buzenkova AV — literature review, data analysis, acquisition and statistical processing of the results, manuscript writing; Grigoryeva ES — flow cytometry, data analysis, interpretation of the results, manuscript writing; Alifanov VV — flow cytometry, manuscript editing; Tashireva LA, Savelieva OE — discussion, manuscript editing; Pudova ES — flow cytometry; Zavyalova MV — manuscript editing; Cherdyntseva NV — study planning and design, discussion; Perelmuter VM — study planning and management, interpretation of the results, manuscript writing.
Compliance with ethical standards: the study was approved by the Ethics Committee of the Cancer Research Institute, Tomsk National Research Medical Center of the Russian Academy of Sciences (protocol No. 8 dated 17 June 2016) and conducted in accordance with Federal Laws of the Russian Federation (No. 152, 323, etc.), the Declaration of Helsinki (1964) and all later amendments and additions that regulate scientific research involving human biomaterial. All subjects submitted the informed consent to participation in the study.
It is difficult to detect the circulating tumor cells (CTCs) being through the epithelial–mesenchymal transition (EMT) terminal phase, since these do not express epithelial markers or show weak expression of those. This hampers assessment of the CTC prognostic potential. It has been shown that the circulating cells (CCs) with the CD45–EpCAM–CK7/8–CD24+N-cadherin‒ phenotype are associated with the risk of metastasis in breast cancer (BC). The study aimed to test CCs based on the side scatter parameters considering the expression of epithelial cell markers and CD11b. CC phenotypes were assessed by flow cytometry within the regions with low (SSClow) and high (SSChigh) side scatter in 11 donors and 20 female patients with BC. All the CD45–EpCAM–CK7/8–CD24+N-cadherin– CCs were represented by the CD11b– and CD11b+ phenotypes found in both SSClow and SSChigh regions. Among eight CD45–mEpCAM–CK7/8–CD24+N-cadherin– CC phenotypes with different variants of co-expression of epithelial markers (E-cadherin, panCK, and icEpCAM) and CD11b found in patients, six showed signs of epithelial nature based on one of the markers, while another two showed no epithelial traits and predominated over other phenotypes (only these two phenotypes were found in donors). The differences in light scattering parameters of the CCs with the same phenotype is one more characteristic, the prognostic value of which remains to be uncovered. The E-cadherin and panCK expression in the absence of mEpCAM and presence of icEpCAM suggest that some CCs are tumor cells in the state of pronounced EMT. CCs showing co-expression of CD11b and epithelial markers can emerge due to hybridization with myeloid cells.
Keywords: breast cancer, flow cytometry, circulating cells, side scatter