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Background. Integrins, as adhesion molecules, play a key role in the interaction of cells with the basal membrane and intercellular matrix. Numerous studies demonstrate evidence of increased expression of integrins on tumor cells in different types of cancer. Thus, β3 and αV integrins are associated with stem-like features of tumor cells, and β4 integrin as α6β4 heterodimer provides anchorage-independent survival of malignant mammary epithelial cells. However, all the described functions of integrins have been investigated exclusively on primary tumor cells. The functional significance and expression pattern of integrins on circulating tumor cells (CTCs) remains unclear. The aim of the study was to evaluate the β3, β4 and αvβ5 integrin expression on CTCs and its association with molecular subtype, stage and lymph node metastasis in breast cancer patients Material and Methods. The study included 22 patients with T1–4N0–3M0 invasive ductal breast carcinoma. Venous blood was taken from patients without neoadjuvant chemotherapy (group 1) and after neoadjuvant chemotherapy (group 2) in the volume of 12 ml into vacuum tubes with EDTA. The expression of CTC integrins including stemness features CD44/CD24, CD133 and ALDH1, and epithelial-mesenchymal transition (EMT) (N-cadherin) was evaluated by flow cytometry. Results. CTCs with β3+β4-αvβ5- and β3-β4+αvβ5+ phenotypes and stemness properties were associated with larger tumor size (T4) in breast cancer patients. The β3 integrin expression was associated with more aggressive molecular subtypes of breast cancer. Administration of neoadjuvant chemotherapy did not affect the expression pattern of β3, β4 and αvβ5 integrins in CTCs. Conclusion. In breast cancer, most CTCs expressed β3, β4 and αvβ5 integrins despite the lack of attachment to the basal membrane and intercellular matrix. The expression of the above integrins on CTCs was associated with breast cancer molecular subtype, stage and lymph node metastasis, and therefore its evaluation can be considered as one of the objectives of liquid biopsy study.

Introduction. Colorectal cancer is a frequently diagnosed disease being in the third place among oncological diseases both in incidence and mortality. Currently, researchers focus on development of more accessible and reliable biomarkers of colorectal cancer to overcome the problems in diagnosis and progression prognosis of this pathology.Aim. To investigate characteristics of microRNA expression in circulating tumor cells (CTC) of patients with colorectal cancer. Materials and methods. The study included blood samples from 299 patients with colon cancer, stages II (T3–4N0M0), III (T1–4N1–2M0) and IV (T1–4N0–2M1). Circulating tumor cells were identified using EpCAM marker detection system. Relative expression of hsa-let-7i-5p, hsa-miR-126-5p, hsa-miR-143-3p, hsa-miR-21-5p, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-92a-3p micro RNA in CTC was measured using polymerase chain reaction.Results. Positive CTC status was observed in 188 (62.9 %) of 299 patients, negative in 111 (37.1 %). In the patient group with pT1–2 tumors, the majority of patients did not have CTC (53.3 %). In other patients with pT1–2 disease, the number of CTC was 1.2 and 4.4 times lower than in patients with pT3 and pT4 disease, respectively. In pT4, 1–3 CTC were found 2.7 and 1.7 times more frequently, 3 CTC 1.4 times more frequently than in pT1–2 and pT3, respectively (p ≤0.05). Presence of metastatic lesions increases the probability of CTC detection by the factor of 2.1: in metastases, >3 CTC were observed 60.1 times more frequently than in M0 (p ≤0.05). Expression of hsa-miR-143-3p and hsa-miR-26a-5p microRNA in CTC of patients with stage III colorectal cancer was respectively 2.5 and 5 times lower than in patients with stage II disease (p

Purpose: to summarize available data on the diagnostic value of various circulating biomarkers for the detection of glioblastoma recurrence.Material and Methods. A literature search was conducted using PubMED ExoCarta and SILVA databases.Results. Glioblastoma multiforme (GBM) is the most common glioma in adults with an unfavorable prognosis. Treatment of tumor recurrence can improve the survival of patients. Neuroimaging is the standard method of diagnosing brain tumor recurrence. However, a neuroimaging method to clearly distinguish between pseudo progression and tumor progression has not been found to date. Current molecular tumor profling relies heavily on tissue resection or biopsy. Tissue profling has several disadvantages in the central nervous system’s tumors, including the challenge associated with invasive biopsy, the heterogeneous nature of many malignancies where a small biopsy can under represent the mutational profle. Liquid biopsy is a promising method in diagnosing malignant tumors. Blood collection is a simple, minimally invasive procedure, but cerebrospinal fuid allows tumor markers to be detected more confdently. However, collection of cerebrospinal fuid is a complex and invasive procedure that can be accompanied by serious complications.Conclusion. Biological fuid markers such as circulating tumor cells, extracellular vesicles, cell-free DNA and cell-free RNA allow for the detection of GMB, determination of molecular genetic features of cancer during response to therapy, and early detection of GBM recurrence.

Influencing the pre-metastatic niche is a very perspective cancer treatment strategy in order of preventing metastases formation. It was found that bone marrow progenitor cells and tumor cells secreting biological compounds are key components in the formation of the pre-metastatic niche. Myeloid suppressor cells (MSCs) are the main type of bone marrow cells in pre-metastatic niches. At the same time, tumor-associated chronic inflammation induces the expression of proinflammatory cytokines triggering myeloid cells differentiation into myeloid suppressor cells. When circulating tumor cells enter the circulatory channel, their interaction with immune cells is observed, which additionally influences the pre-metastatic site preparation. Studies have shown that the entire spectrum of immune cells is capable of influencing the metastasis formation by circulating tumor cells. The epithelialmesenchymal transition with the tumor cell transporting form appearence was found to be related to the function of the ZEB1 protein. Its activity is regulated by numerous signaling mechanisms at the transcriptional level, including TGFβ, Wnt and Notch. This initiates epithelial-mesenchymal transition of breast cancer cells. Zhang Z.et al. proved that CDK4/6 blocking leads ZEB1 protein stability decreasing, preventing metastasis in breast cancer in vitro and in vivo. Moreover, USP51 deubiquitinase has been identified as a target of cyclin-dependent 4/6 kinases. At the molecular level, CDK4/6 phosphorylate and activates USP51, which then influences ZEB1 deubiquitination and stabilization. A positive correlation was demonstrated between the expression of p-RB (an indicator of CDK4/6 activity), p-USP51 and ZEB1 in breast cancer samples. Thus, the CDK4/6-USP51-ZEB1 axis may play a key role in the metastasis of breast cancer. In breast cancer cells, inhibition of CDK4/6 was shown to increase the expression of E-cadherin but decrease the expression of mesenchymal markers, reducing the migratory ability and invasiveness of breast cancer cell lines. This biological effect may also explain the clinical efficacy of the CDK4/6 inhibitor Abemaciclib in early-line therapy of metastatic breast cancer as well as in adjuvant combination hormone therapy for the prevention of metastatic lesions in patients at high risk of recurrence and progression in the MONARCH E study. Besides, there were no response predictors evaluated in trials investigating CDK4/6 in breast cancer treatment and it is unknown if there any differences in treatment response according to the metastatic site. The clinical cases demonstrate abemaciclib clinical efficacy in metastatic breast cancer treatment regardless of metastatic site.

Introduction. Ability of circulating tumor cells (CTC) initiate metastases in distant sites is associated primarily with their resistance to apoptosis which allows them to retain viability in the blood. Knowledge of phenotypical signs associated with this ability would allow to predict the risk of metastases and optimize adjuvant therapy.Aim. To examine signs of apoptosis in CTC populations with various phenotypical characteristics.Materials and methods. The study included 58 patients with invasive breast carcinoma of unspecified type, stages T1–4N0–3M0. Cell concentrates extracted from patients’ whole blood were stained with an antibody cocktail against CK7 / 8, CD45, EpCAM, CD44, CD24, CD133, ALDH, N-cadherin which allowed to identify CTC with signs of stemness and epithelial-mesenchymal transition. Annexin V and 7‑amino-actinomycin D staining was used for evaluation of apoptosis stage in CTC populations.Results. Circulating tumor cells are characterized by heterogeneity in respect to signs of stemness and epithelial-mesenchymal transition and presence of early and late signs of apoptosis and necrosis. CTC phenotypes including co-expression of epithelial marker CK7 / 8 and stemness marker CD133 (but not CD44) are characterized by absence of signs of apoptosis. Co-expression of CK7 / 8 and CD133 in CTC with stemness markers CD44+ / C D24– is associated with development of early but not late signs of apoptosis and necrosis. Circulating tumor cells without co-expression of CK7 / 8 and CD133 could have both early and late signs of apoptosis and necrosis. Circulating tumor cells phenotypes with signs of early apoptosis expressing CD133 remain in blood after non-adjuvant chemotherapy opposed to CTC without CD133 expression.Conclusion. There are CTC phenotypical signs associated with stemness and epithelial-mesenchymal transition and linked to apoptosis resistance or sensitivity.

Background. Distant metastasis remains the main cause of death for patients, despite some successes in the diagnosis and treatment of solid tumors. This demonstrates the significance to study the methods for the timely detection of tumor micrometastases and to find the role of tumor cells preserved in peripheral blood or disseminated tumor cells in bone marrow after the end of the treatment. Aim. To generalize the data concerning the significance and methodology of the determining circulating tumor cells in peripheral blood and disseminated tumor cells in bone marrow in case of different types of cancer. Materials and methods. Foreign and domestic articles concerning this topic, published in eLibrary, PubMed, Medline, Scopus databases have been analyzed. Results. The literary review deals with the modern methods of determination, we have analyzed the importance of identification of circulating and disseminated tumor cells as the clinical predictors for specific nosological types of cancer. Conclusion. The enumeration of circulating and disseminated tumor cells could be a new generation of screening and evaluation of the efficacy of cancer therapy. However, it is necessary and up to date to develop the methodology of their quantitative and qualitative identification and to find the methods for evaluating functional activity.

Introduction. Serum chemokines are inflammatory mediators, which role is shown in the occurrence and progression of a number of malignant tumors. Produced by white blood cells, stem cells, tumor and endothelial cells, chemokines control their movement and positioning. Chronic inflammation underlies the progression of ovarian cancer (OC ). This increases the likelihood of chemokines stimulating or blocking tumor progression.The aim of the study was to evaluate the relationship between the blood levels of inflammatory cytokines in blood and the number of circulating tumor cells (CTC s) with the response to standard chemotherapy (CT ) in patients with cancer.Material and Methods. In patients with primary OC before and after 2–4 courses of chemotherapy and in patients with benign ovarian tumors (as a control), serum levels of CCL 2, CCL 3, CCL 4, CXCL 8 and CX3CL 1 were evaluated by multiplex xMAP analysis. The amount of CTC s (population CD 45-/ Epcam+/CK+) was determined using a flow cytometer. Patients with ovarian cancer were divided into 3 groups according to the platinum sensitivity criterion of GC JG 4th, and progression-free interval (PFI) was determined. Results. It was found that the levels of CCL 2, CCL 3, CCL 4, CXCL 8, and CX3CL 1 in case of OC did not significantly differ from that in the control, strongly negatively correlated with age (except for the CCL 2 level). CT significantly increased the level of CCL 2 in the group of refractory OC ; of CCL 3 – in the group of sensitive OC , of CCL 4 – in the groups of resistant and sensitive OC , and C XCL 8 level increased in the groups with resistant and sensitive OC and decreased in the group of refractory OC . The number of CTC s in patients with OC was significantly higher than in the control. After CT , a decrease in the amount of CTC s strongly and significantly correlated with a decrease in the level of CX3CL 1 in the groups of refractory andsensitive OC . The maximum PFI occurred with an increase in serum levels of CCL 3, CXCL 8, a decrease in CCL 4 and a constant level of CX3CL 1.Conclusion. Thus, no significant differences in the levels of CCL 2, CCL 3, CCL 4, and IL -8 between patients with OC and control groups were found. The levels of chemokines studied and the amount of CTC s differed in the groups divided by the tumor sensitivity to CT . We observed significant correlations between the amount of CTC s and the level of CX3CL 1 in the group of platinumsensitive OC .

Background. The search and use of new molecular prognostic and predictive markers of efficacy detected by liquid biopsy are aimed at understanding the biology of Carcinomas of Unknown Primary (CUP) and improving results of patient treatment. The review is devoted to advances in scientific and clinical research on this issue.Materials and methods. In order to assess the current state of the problem, a search and analysis of relevant data of the scientific databases PubMed, Medline, RISC was carried out.Results. The scientific rationale for the use of liquid biopsy in clinical practice to improve the treatment of cancer patients with CUP is presented. The results of the use of modern approaches to the analysis of fluid biopsy samples in CUP are presented. In particular, the features of using circulating free DNA, circulating tumor DNA, circulating tumor cells in the analysis are considered. The modern possibilities of determining tissue specificity using liquid biopsy in CUP are discussed. The prospects for the development of liquid biopsy for improving diagnostics, determining the prognosis of the disease, and choosing a strategy for treating CUP and the treatment monitoring have been determined.Conclusion. A review of the literature confirms that modern methods of molecular profiling of tumor cells obtained both as a result of liquid biopsy and tissue biopsies will make a significant contribution to the determination of tissue specificity, molecular characteristics of CUP for a personalized approach in order to improve strategies and treatment outcomes for patients with CUP.

Nowadays, circulating tumor cells (CTCs) are considered to be one of the most important mechanisms of tumor dissemination. Detection of CTCs in patients` blood is estimated as a prognostic factor for various tumors including colorectal cancer (CRC). However, CTCs may also be a source of tumor antigens capable of inducing both immune response and tolerance and participating in «immmunoediting». The aim of the study was to assess the parameters of cell-mediated immunity in patients with stage IIIV CRC with respect to the presence or absence of CTCs. Materials and Methods. We studied the parameters of cell-mediated immunity in 60 patients with stage IIIV CRC with respect to the presence or absence of CTCs. Before treatment, we evaluated the CTC levels in patients’ blood using CellSearch System™ and lymphocyte subsets: Т-В-NК-cells, T-reg, CD4+ and CD8+ expressing markers of activation (CD69+, СD38+, CD25+, HLA-DR+ СD95+); naïve and memory T-lymphocytes (CD45RA-/CD45RO+); NК-cells expressing CD335, perforin and granzyme B using flow cytometry (FACSCantoII, BD). Results. A difference in the immunologic parameters between patients with CTCs and without CTCs depending on the stage of CRC was found. In СTС-positive patients with locally-advanced CRC, increase in the parameters of innate immunity (CD335+ NK-cells, neutrophils` respiratory burst) and activated Th (CD38+, CD25+, HLA-DR+) was found, while in СTС-positive patients with generalized CRC, suppression of cytotoxic lymphocytes of both innate (CD56/16+) and adaptive (CD8+CD25+) immunity was observed, which is, apparently, an unfavorable prognostic factor. Conclusion. The presence of CTC in CRC patients is accompanied by some immunologic changes rather stimulating Thlink and innate immunity factors in II-III stages and doubtlessly suppressive in patients with IV stage.

Background. Breast cancer (BC) is the most common type of malignant neoplasm among women, with a high rate of metastasis. Early non-invasive diagnosis is required to increase the effectiveness of anticancer therapy.Objective: to determine the content of circulating tumor cells (CTCs) and their derivatives in the peripheral blood using the MDA-231 aptamer, compare the results obtained with the clinical and molecular characteristics of BC.Materials and methods. The study included 22 patients with BC. Detection of CTCs and circulating tumor microemboli was carried out in 3.5 ml of the blood of BC patients with the help of the MDA-231 aptamer which is affine for breast cancer cells, labeled with the fluorescent Cy3 dye. The count of CTCs in the blood samples was performed using fluorescent and laser scanning microscopy.Results and conclusions. The content of CTCs and circulating tumor microemboli in the peripheral blood of patients with BC of various molecular subtypes was analyzed using the MDA-231 aptamer. The relationship between the number of CTCs and the molecular biological subtype was revealed. The obtained results show the possible prognostic value of CTCs use for monitoring effectiveness of anticancer therapy and control of recurrence of BC.

Cervical cancer is one of the most common cancers among women of reproductive age. The cytological screening is not always effective and appropriate, therefore the search for new predictive markers of the cervical cancer are of great importance. there are no biomarkers for monitoring patients previously treated for cervical cancer. liquid biopsy is a new option of personalized approach to the detection and monitoring of cervical cancer. it is a set of methods for determining the derivatives of a tumor in biological media, most often in the blood: circulating tumor cells, circulating dNa, RNa, exosomes, etc.The purpose of the studywas to analyze data on the role of liquid biopsy in the diagnosis and monitoring of cervical cancer.Material and methods. We analyzed publications available from pubmed, Elibrary over the past 10 years.Results. Circulating tumor cells, circulating tumor dNa and exosomes are the most studied cancer non-invasive biomarkers. these circulating biomarkers play a key role in the understanding of cervical carcenogenesis, chemo-and radioresistance. currently, liquid biopsy is considered as a promising modern method for the detection and monitoring of cervical cancer. the diagnostic efficiency of this method is good, so it can be used for cervcal cancer screening. However, such statements require further research in this direction. in addition, given the emerging information on the molecular carcinogenesis of cervical cancer, liquid biopsy can also be used as a basis for the development of targeted therapy for locally advanced and generalized cervical cancer.Conclusion. Liquid biopsy is the non-invasive method of cervical cancer monitoring.

The aim of this study was to assess CXCR4 expression in different subsets of CTCs and single (detached) breast cancer cells.Materials and methods. Thirty five patients with invasive breast carcinoma of no specialtype (IC NST) (T1-4N0-2M0), between 29 and 69 years of age were included in this study. Different subsets of CTCs with CXCR4 expression were evaluated by flow cytometry. A confocal microscopy was used to assess CXCR4 expression in different subsets of single (detached) cancer cells in breast tissue.Results. The CXCR4 was expressed in CTCs without stem-like and EMT phenotype, in CTCs with EMT but not stem markers and in stem-like CTCs without EMT features. In all blood samples, the CXCR4 expression in CTCs with stem-like and EMT phenotype was absent. In breast tumor the CXCR4 was expressed in the non stemlike single (detached) breast cancer cells with EMT features, in the single (detached) breast cancer cells with stem and EMT features. In all tumor samples the stem-like or non stem-like single (detached) breast cancer cells without EMT features were absent.Conclusions. Different subsets of the CTCs exhibited CXCR4. The CXCR4 expression did not depend on the presence or absence of stem or/and EMT features in tumor cells. We showed that some subsets of single (detached) breast cancer cells in the primary tumor were characterized by the ability to express CXCR4 and may be a source of the respective CTC subsets.

Breast cancer is the most common female oncopathology in the world. The main cause of mortality in patients - metastases. The current state of medicine allows curing the primary tumor, but often is not effective enough to control the development of metastatic disease. Of great importance in the implementation of metastatic cascade plays circulation and survival of tumor cells in the peripheral blood. In this article we have tried to summarize and present the data in the literature on the study of circulating tumor cells in patients with breast cancer.

In 40 patients with newly diagnosed colorectal cancer identify circulating tumor cells the Veridex CellSearch system. In the tissue of the tumor was determined by the presence of KRAS mutations for Real-Time qPCR technique. Was found a connection of an elevated number of CTCS in the venous blood with the presence of tumor mutations in the KRAS gene. In patients with high levels of CTCS and the presence of mutations, earlier developed distant metastases.

The paper covers the most important topographic-anatomic and histological characteristics of the vagina, gives general information on the epidemiology of primary vaginal cancer (VC), on the rules of its staging, describes the frequently encountered morphological types of this tumor, and indicates the properties and features of the course of neuroendocrine tumors (NETs), including the most common manifestations of carcinoid syndrome and the symptoms of tumor hypersecretion of a number of biologically active substances. It also indicates the most common sites of NETs, the rules of their classification according to the grade and mitotic activity of tumor cells, as well as major methods for biochemical and radiological diagnosis. The immunohistochemical NET markers that can diagnose VC, their physiological role, sensitivity, and specificity, as well as the possibility of determining them in the body’s biological environments in order to increase physicians’ awareness are described in detail and the main aspects of tumor immunohistochemical staining are denoted. The role of circulating tumor cells as a promising method to specify the extent of NETs is defined. The paper gives epidemiological information on the development of vaginal NET and summarizes the available data by their clinical course and treatment results. It provides current guidelines on the required treatment volume and specifies the role of surgical intervention and the necessary volume if the intervention is expedient. It sets forth the up-to-date guidelines for chemotherapy for small cell tumors, as well as VC, which includes the specific features of managing patients with tumor recurrences depending on the time of their development.