Stem properties allow circulating tumor cells (CTCs) to survive in the bloodstream and initiate cancer progression. We aimed to assess the numbers of stem-like CTCs in patients with ovarian cancer (OC) before treatment and during first-line chemotherapy (CT). Flow cytometry was performed (Cytoflex S (Beckman Coulter, CA, USA)) using antibodies against CD45; epithelial markers EpCAM and cytokeratin (CK) 8,18; mesenchymal vimentin (vim); and stem-like CD44, CD133 and ALDH. This study included 38 stage I–IV OC patients (median age 66 (Q1–Q3 53–70)). The CK+vim- counts were higher (p = 0.012) and the CD133+ALDHhigh counts were lower (p = 0.010) before treatment in the neoadjuvant CT group than in the adjuvant group. The patients with ascites had more CK+vim- cells before treatment (p = 0.009) and less EpCAM-vim+ cells during treatment (p = 0.018) than the patients without ascites. All the CTC counts did not differ significantly in paired samples. Correlations were found between the CK-vim+ and CD133+ALDHhigh (r = 0.505, p = 0.027) and EpCAM-vim+ and ALDHhigh (r = 0.597, p = 0.004) cells before but not during treatment. Multivariate Cox regression analysis showed that progression-free survival was longer with the presence of surgical treatment (HR 0.06 95% CI 0.01–0.48, p = 0.009) and fewer CD133+ALDHveryhigh cells (HR 1.06 95% CI 1.02–1.12, p = 0.010). Thus, CD133+ALDH+ CTCs have the greatest prognostic potential in OC among the phenotypes studied.

Ovarian cancer (OC) is able to develop implantation metastases in the abdominal cavity. Ascites is potentially useful for evaluating cancer features. The aim of the study was to assess the content of stem-like tumor cells and inflammatory mediators in ascites of OC. The prospective study included 11 patients with primary OC having ascites, 8 patients with benign ovarian tumors having ascites and 22 healthy women. In ascitic fluid obtained by laparocentesis, the populations of tumor stem-like cells were determined on a Cytoflex S` flow cytometer (Beckman Coulter, USA) and CytExpert Software using monoclonal antibodies to CD45, CD44 and CD133. The cytokine profiles of ascitic fluid and blood serum (IL-1β, IL-18, IL-4, IL-10 and VEGF) were assessed by ELISA. Stem-like cells were found in all samples. 5 cell populations were evaluated. The number of cells expressing both markers: CD44 + and CD133+, was the lowest. The highest, about 32%, was the number of CD44+ cells. The number of cells CD45-CD44+CD133- in ascites strongly positively correlated with the content of IL-10 in ascites, and the numbers of CD45-CD133+ and CD45-CD44-CD133+ - with the level of VEGF in blood serum. No correlations were found between the numbers of stem-like cells and the disease stage or the level of CA125 in blood. The combination of IL-4 and IL-10 in ascites had the greatest significance in predicting the disease stage. These results suggest a relationship between the levels of VEGF, IL-10, and cancer stem cells in the OC ascites. Stem-like cells in OC ascites are heterogeneous and are present even at an early stage of the disease. It seems promising to study cell populations and cytokine profile of ascites together, to assess the biomarker potential of their combination.

Background: Circulating Tumor Cells (CTCs) are a potential source of metastases and relapses. The data on molecular characteristics of Ovarian Cancer (OC) CTCs are limited. Objective: This study aims to assess the TGFβ, CXCL2, VEGFA and ERCC1 expressions in two OC CTC subpopulations before and during chemotherapy (CT), and their relation to clinical characteristics. Methods: Two CTCs subpopulations (EpCAM+CK18+E-cadherin+; EpCAM+CK18+Vimentin+) were enriched using immunomagnetic separation before treatment and after 3 cycles of platinumcontaining CT. The expression of mRNA was assessed using RT-qPCR. Results: The study included 31 I-IV stage OC patients. During CT, TGFβ levels increased in both fractions (p=0.054) compared with the initial levels. ERCC1 expression in E-cadherin+ CTCs was higher during neoadjuvant than adjuvant CT (p=0.004). CXCL2 level in E-cadherin+ CTCs increased (p=0.038) during neoadjuvant CT compared with the initial. TGF-β expression in vimentin+ CTCs during CT was negatively correlated to disease stage (p=0.003). Principal component analysis before CT revealed a component combining VEGFA, TGFβ, CXCL2, and a component with ERCC1 and VEGFA; during CT, component 1 contained ERCC1 and VEGFA, and component 2 - TGFβ and CXCL2 in both fractions. Increased ERCC1 expression in E-cadherin+ CTCs during CT was associated with decreased Progression-Free Survival (PFS) (HR 1.11 (95% CI 1.03-1.21, p=0.009) in multivariate analysis. Conclusion: EpCAM+ OC CTCs are phenotypically heterogeneous, which may reflect variability in their metastatic potential. CT changes the molecular characteristics of CTCs. Expression of TGFβ in EpCAM+ CTCs increases during CT. High ERCC1 expression in EpCAM+CK18+E-cadherin+ CTCs during CT is associated with decreased PFS in OC.

Фенотип циркулирующих нейтрофилов на разных стадиях неоплазии шейки матки

Summary. Circulating tumor cells (CTCs) are a potential source of tumor progression. Systemic tumor-associated inflammation can influence the cancer prognosis both independently and by changing the characteristics of CTCs. Data on the role of proinflammatory cytokines and CTCs in the ovarian cancer (OC) chemosensitivity are few and contradictory. Aim of the study. To assess the IL-17A, IL-18 levels and the number of CTCs in primary patients with OC before treatment and after 3 courses of standard platinum-containing chemotherapy, and the possibility of using these cytokines as a marker of the presence of CTCs. Materials and methods. The study included 72 patients with OC. The comparison group included 16 patients with benign ovarian tumors, the control group included 20 healthy women. The number of CTCs (CD45-/EpCam+/CK+) was determined immunofluorometrically before and after 3 courses of chemotherapy. The content of cytokines was assessed by ELISA. Statistica 13.0, jamovi 1.6.5.0 were used for data processing. Results. The level of IL-17A in blood in the benign tumors group was increased in comparison with OC (р=0.012) and control (р=0.042). The content of IL-17A increased over time in the subgroup of adjuvant chemotherapy (р=0.017). During treatment, IL-17A was higher among patients with platinum-sensitive tumors than with non-sensitive ones (р=0.054). High levels of IL-18 before treatment were associated with the development of platinum-refractory relapse (р=0.014). IL-18 levels before (р=0.027) and during treatment (p=0.052) were lower in patients with cytoreductive surgery in first line of treatment. Progression-free survival (PFS) (p = 0.012) and overall survival (OS) (p = 0.030) were lower in the cluster with high IL-18 and pretreatment leukocyte counts than in the cluster with low scores. Higher quantity of the pretreatment CTCs was associated with longer PFS in a multivariate analysis (HR 0.82 95% CI 0.69–0.98, р=0.028). The number of CTCs before treatment greater than 5 was associated with a decrease in OS (HR 1.31, 95% CI 0.98–1.73, р=0.064). Conclusions. A high level of IL-17A in the blood during treatment is associated with more favorable clinical characteristics of OC, while a high level of IL-18 in the blood before treatment is associated with less favorable characteristics and a worse prognosis. The number of CTCs in patients with OC is not related to the blood levels of IL-17A and IL-18. A greater number of CTCs before treatment in OC is associated with an increase in PFS, but a decrease in overall survival. Циркулирующие опухолевые клетки (ЦОК) — потенциальный источник опухолевой прогрессии. Системное опухоль-ассоциированное воспаление способно влиять на прогноз заболевания и изменять характеристики ЦОК. Цель. Оценка уровней IL-17A, IL-18 и количества ЦОК у первичных пациенток с раком яичников (РЯ) до лечения и после 3-х курсов платиносодержащей химиотерапии (ХТ) и их связи с ЦОК. Материалы и методы. В исследование включены 72 пациентки с РЯ. Группа сравнения включала 16 пациенток с доброкачественными опухолями яичников, контроль — 20 здоровых женщин. Количество ЦОК определяли иммунофлуориметрически (CD45-/EpCam+/CK+). Содержание цитокинов оценивали методом ИФА. Для статистической обработки использовали Statistica 13.0, jamovi 1.6.5.0. Результаты. Содержание IL-17A в крови при доброкачественных опухолях было повышено в сравнении с РЯ (р=0,012) и контролем (р=0,042). Уровень IL-17A повышался в динамике при адъювантной ХТ (р=0,017). В ходе лечения показатель IL-17А был выше среди пациенток с платиночувствительными опухолями, в сравнении с нечувствительными (р=0,054). Высокое содержание IL-18 до лечения было связано с развитием платинорефрактерного рецидива (р=0,014). Уровни IL-18 до лечения (р=0,027) были ниже у пациенток с циторедукцией в первой линии. Выживаемость без прогрессирования (ВБП) (р=0,012) и общая выживаемость (ОВ) (р=0,030) были ниже в кластере с высокими IL-18 и числом лейкоцитов до лечения, чем в кластере с низкими показателями. Большее число ЦОК до лечения было ассоциировано с более длительной ВБП (HR 0,82, 95% ДИ 0,69–0,98, р=0,028). Число ЦОК до лечения более 5 было связано со снижением ОВ (ОР 1,31, 95% ДИ 0,98–1,73, р=0,064). Выводы. Высокий уровень IL-17A в крови ассоциирован с более благоприятными клиническими характеристиками РЯ, а высокий уровень IL-18 в крови до лечения — с менее благоприятными характеристиками и худшим прогнозом. Количество ЦОК не связано с содержанием в крови IL-17A и IL-18. Большее число ЦОК до...