e12581 Background: Despite the efficacy of anthracyclines in breast cancer (BC) treatment, their use is limited by cardiotoxicity and resistance mechanisms. Secondary plant metabolites offer a reservoir for novel cytostatics. We investigated the in vitro activity of a novel indole alkaloid (P1), isolated from Petasites hybridus, and its interaction with doxorubicin (DOX) across clinically relevant BC subtypes. Methods: Cytotoxicity was evaluated using the MTT assay (48h exposure) in three human BC cell lines: MDA-MB-453 (HER2+, ER-/PR-), BT-474 (Luminal B/HER2+), and BT-20 (Triple-Negative BC), compared to primary normal skin fibroblasts as a non-malignant control. Half-maximal inhibitory concentrations (IC50) were determined. Drug interaction landscapes were analyzed using the Zero Interaction Potency (ZIP) model via SynergyFinderPlus software. Interaction was classified based on Synergy Scores (SS): >10 (synergy), -10 to 10 (additive), <-10 (antagonism). Results: P1 exerted dose-dependent antiproliferative effects in all BC lines. IC50 values were 31.74±3.8 µM for MDA-MB-453, 39.70±2.4 µM for BT-20, and 49.23±5.2 µM for BT-474. Notably, P1 demonstrated selective toxicity towards cancer cells; viability of all BC lines was significantly lower than that of normal fibroblasts at P1 concentrations >22 µM (p<0.05), indicating a favorable therapeutic window. DOX showed expected high potency (IC50 0.51–10.4 µM) but exhibited significant antagonism when combined with P1 in HER2+ cell lines: mean SS was -14.98 for BT-474 and -8.23 for MDA-MB-453 (reaching <-17 at high doses). Conversely, in the TNBC line BT-20, the combination was additive (mean SS -2.66), suggesting no interference with DOX efficacy. Conclusions: The novel indole alkaloid P1 demonstrates selective cytostatic activity against breast cancer cells sparing normal fibroblasts. The drug interaction profile is highly subtype-dependent: while P1 antagonizes DOX in HER2+ models, likely due to cell cycle interference, it shows additive potential in triple-negative breast cancer. These findings position P1 as a promising lead compound for TNBC therapy requiring further mechanistic elucidation. Comparative IC50 and interaction scores. Cell Line Molecular Subtype P1 IC50 (µM) Interaction with Doxorubicin (Mean Synergy Score) Interpretation MDA-MB-453 HER2+ 31.74 ± 3.8 -8.23 Antagonism BT-474 Luminal B / HER2+ 49.23 ± 5.2 -14.98 Antagonism BT-20 Triple-Negative 39.70 ± 2.4 -2.66 Additive

e23540 Background: Cellular bioenergetic profiles are critical determinants of tumor biology and can vary significantly between different malignancies. Characterizing the metabolic dependencies of primary sarcoma cultures is essential for understanding their pathophysiology and potential therapeutic vulnerabilities. This study aimed to define the energetic phenotypes of primary sarcoma cell cultures by assessing their metabolic response to glucose. Methods: Five primary sarcoma cell cultures were established from patient samples via enzymatic dissociation with collagenase I and cultured in DMEM medium supplemented with 10% FBS, ITS, and NEAA. Metabolic profiling of adherent cells was performed using a Seahorse XFp analyzer. Assays included the mitochondrial stress test, FCCP titration, and the glycolytic stress test according to the manufacturer's protocols, utilizing media optimized for specific energetic substrates to accurately measure cellular respiration and glycolysis. Results: Analysis of the respiratory response to glucose addition revealed two distinct metabolic phenotypes. Group 1 (three cultures) exhibited a low mitochondrial ATP production rate (<5% of total respiration linked to ATP synthesis) but high reserve respiratory capacity (>150% of baseline) and glycolytic reserve (>1600% of baseline). As expected, these cultures increased oxygen consumption upon glucose addition, proportional to their high reserve capacities. In contrast, Group 2 (two cultures) demonstrated a high mitochondrial ATP production rate (>10%) but low reserve respiratory capacity (<90%) and low glycolytic reserve (<700%). These cultures decreased oxygen consumption following glucose addition, corresponding to a reduction in their spare metabolic capacities. Conclusions: Primary sarcoma cultures display two distinct energetic phenotypes: a "high-reserve" phenotype with low baseline mitochondrial ATP production but substantial capacity to upregulate both oxidative phosphorylation and glycolysis in response to glucose; and a "low-reserve" phenotype with high baseline mitochondrial ATP dependence but limited ability to further enhance metabolism with increased glucose. This phenotypic stratification may reflect underlying biological heterogeneity in sarcomas and could inform strategies for metabolic targeting.

2658 Background: Promising oncolytic viruses are typically evaluated based on their direct cytotoxic effects on cancer cells. However, the therapeutic benefit of virotherapy may also stem from its immunomodulatory actions, particularly through influencing immune checkpoint molecules like PD-1 and PD-L1. This study aimed to investigate the effect of unclassified apathogenic rotavirus strains RVK100 and RVK228 on the expression of PD-1 and PD-L1 on T-cells derived from the peripheral blood of patients with breast cancer. Methods: Mononuclear cells of peripheral blood were isolated on a ficoll gradient, cultured in RPMI 1640 (Gibco, USA) without serum at 37 °C, 5.0% CO2 in 4 variants of the experiment: 1) negative control without viruses; 2) positive control of activation with the addition of PHA; 3) experience with the addition of 107 particles per 1 ml of the RVK100 strain; 4) experience with the addition of 107 particles per 1 ml of the RVK100 strain RVK228. After 24 and 72 hours of cultivation, the expression of PD-1 (CD279) and PD-L1 (CD274) was determined on T cells by flow cytometry. The study used antibodies conjugated with fluorochromes: anti-CD4 (PE), anti-CD8 (APC Cy7), anti-CD279 (FITC), anti-CD274 (PerCP-Cy5–5) (Becton Dickinson, USA). Results: After 24 hours, we observed increases in PD-1 expression on CD4+ (PHA—40.5%, RVK100—42.3%, RVK228—37.5%; vs. control—18.1%) and CD8+ cells (PHA—41.7%, RVK100—46.4%, RVK228—42.6%; vs. control—27.7%). Similarly, PD-L1 expression rose on CD4+ cells (RVK100—67.0%, RVK228—58.6%, PHA—75.1%; vs. control—44.8%) and CD8+ cells (RVK100—63.4%, RVK228—58.4%, PHA—52.8%; vs. control—46.2%). At 72 hours, PD-1 levels decreased significantly in CD4+ cells exposed to RVK100 (from 42.3% to 21.6%) and CD8+ cells (from 46.4% to 17.4%). Conversely, PD-L1 expression increased across all groups on CD8+ cells, reaching 67–79%, while minimal change occurred on CD4+ cells except for a minor decline in the RVK100 group. Conclusions: Both strains, like the non-specific T-mitogen PHA, caused the stimulation of the expression of immune checkpoint receptors PD-1 and PD-L1 on T-helpers and CTL after 24 hours of cultivation. After 72 hours of cultivation, RVK100, unlike RVK228, was revealed ability to reduce the expression of PD-1 on these cells.

e14056 Background: Brain tumors present a genuine challenge for medicine due to their anatomical features, high resistance to existing chemotherapeutic drugs, a strongly immunosuppressive environment, and the presence of the blood-brain barrier. The research and development of new anti-tumor drugs remains an important task. Tropolone derivatives represent a promising class of organic compounds that could serve as new pharmaceutical anti-tumor agents. There is currently a growing interest in tropolone alkaloids, as many of them possess a broad spectrum of pharmacological activity. The aim of the study was to investigate the cytotoxic effect of a new tropolone derivative 2-(1,1-dimethyl-1H-benzo[e]indoline-2-yl)-5,6,7-trichloro-1,3-tropolone (JO-122(2)) on primary glioma cell cultures in an in vitro experiment. Methods: The tumor material for primary cell cultures was obtained from a patient who was diagnosed with grade 4 glioblastoma on the background of anaplastic astrocytoma (Culture 1) and a patient with histologically verified grade 4 giant cell glioblastoma (Culture 2) who were treated at the Department of Neuro-Oncology of the National Medical Research Centre for Oncology of the Ministry of Health of the Russian Federation. Cell cultures were cultured on DMEM medium (Gibco, USA) with the addition of 10% FBS (Hyclone, USA). The cells were planted in a 96-well plate (Eppendorf, Germany) in the amount of 5,000 cells in 100 µl of medium per well and incubated for 24 hours at a temperature of 37 ° C in an atmosphere containing 5.0% CO2. After 24 hours, the culture medium was replaced with a medium containing JO-122(2) in a series of double dilutions. The maximum amount of JO-122(2) was 24 μmol. The MTT test was performed according to the standard procedure, the exposure time was 24, 48 and 72 hours. The optical density was determined at a wavelength of 540 nm using a tablet reader. The inhibition index (IC50) was defined as the percentage of surviving cells after exposure to JO-122(2) from the control. Results: The results of the in vitro cytotoxic effect study showed that the IC50 for Culture 1 after incubation with the addition of JO-122(2) for 24 hours was 5.3668 μmol, 48 hours - 3.9358 μmol, 72 hours - 1.5418 μmol. For Culture 2, the IC50 after incubation for 24 hours was 2.9146 μmol, 48 hours - 2.1581 μmol, 72 hours - 1.1187 μmol. Conclusions: Our results indicate that JO-122(2) has a pronounced cytotoxic effect on primary glioma cell cultures. It is noted that an increase in the duration of incubation contributes to a decrease in cell viability.

e19114 Background: T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy that accounts for 25% of all adult cases of T-ALL. Currently, the frequency of disease recurrence and resistance to therapy remains quite high, so there is an urgent need to improve therapy by testing new compounds that may potentially have antitumor activity. Promising compounds include berberine, whose high antitumor activity we previously confirmed on glioblastomas, and its derivatives. The aim of the study was to evaluate the cytotoxic effect of berberine derivatives on Jurkat cell culture in an in vitro experiment. Methods: Jurkat cell culture cells were cultured in a complete RPMI1640 nutrient medium without phenolic red (Gibco, USA) with the addition of 10% FBS (Hyclone, USA), 1% glutamine (Biolot, Russia). A total of 11 compounds were tested: berberine and its derivatives ZbRNN, Rub, NAPh, Rub HCl, 12NO2, EPOX, Mag-2, Mag-1, AkR(1) Fuz, 21A. Dose-response curves were constructed for each compound, measuring the level of metabolically active cells by colorimetric method using tetrazolium salt XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide). The exposure time was 72 hours. Viability was determined as the ratio of the number of living cells in the experimental wells to the control wells, expressed as a percentage. Based on the data obtained, dose-response curves were constructed and IC50 values were determined using the dcr package of the R programming language. Results: Compounds MAG-1, MAG-2, 21A, ZbRNN, Rub, 12NO2, EPOX, and NAPh demonstrated high activity (IC50 below 1000 nmol/L) against the Jurkat cell line. The IC50 value for the studied substances was: 236.4±33.5 nmol/L, 31.8±9.1 nmol/L, 126.3±27.2 nmol/L, 103.8±13.3 nmol/L, 136.4±18.7 nmol/L, 308.4±39.3 nmol/L, 455.7±54.1 nmol/L and 789.5±26.6 nmol/l, respectively (p<0.05). For compounds Rub HCl and AkR(1) Fuz, the IC50 value exceeded 1000 nmol/l and amounted to 1756.2±99.1 nmol/l and 1435.7±112.8 nmol/L (p<0.05). For berberine, the IC50 value was 447.6±46.1 nmol/L. Conclusions: The results obtained indicate the pronounced cytostatic properties of a number of berberine derivatives, of particular interest are compounds that are close to and superior to berberine in cytostatic properties.

Importance: Cutaneous melanoma is regarded as the most lethal form of skin cancer. According to the literature, issues associated with accurate prognosis and early detection of relapses hinder effective treatment of melanomas. Over the past 20 years, microRNAs have been shown to modulate various biological processes. Disruption of microRNA expression and activity can contribute to the risk of cancer development, including cutaneous melanoma. Therefore, we decided to study some microRNAs as biomarkers for future use in cutaneous melanoma diagnosis.Objective: To analyze the expression of microRNA-205, microRNA-199a-3p, microRNA-146a-5p in tumor tissue of patients diagnosed with cutaneous melanoma.Materials and methods: We quantified the expression of microRNA-205, microRNA-199a-3p, microRNA-146a-5p using the realtime polymerase chain reaction in biopsy specimens from patients with melanoma (n=14). For all the specimens, the cycle threshold (Ct) value was 0.1 (qPCRsoft 3.0, Analytik Jena, Germany). Statistical analysis was performed using Statistica 7.0 (StatSoft, USA). Furthermore, we conducted a bioinformatics search for microRNA target genes using DIANA-mirPath v.3.0 (DIANA-Lab, Greece) and constructed gene interaction networks using STRING (https://string-db.org/).Results: We detected 2 of 3 microRNAs in the group of patients with cutaneous melanoma: microRNA-205 and microRNA-146a-5p. In both the control group and the group of patients with melanoma, 1 of 3 microRNAs, namely miR-146a-5p, was found. The mean ∆Ct value of microRNA-146a-5p was 3.81±2.03 in the control group and 2.26±1.34 in the patients with melanoma. Due to the wide range of values in the control group and the group of patients with cutaneous melanoma, microRNA-146a-5p cannot be considered an informative biomarker. At the same time, we found high expression of microRNA-205 in the group of patients with cutaneous melanoma.Conclusions: microRNA-205 was found to be valuable in melanoma diagnosis. However, further studies with larger sample size and additional stratification by histopathological features are needed.

Мета-анализ был направлен на оценку связи между полиморфизмами rs2107425, rs2839698, rs217727, rs3741219 гена H19 и риском развития рака молочной железы. Поиск публикаций проводили в базах данных Google Scholar и PubMed за последние 13 лет. Ассоциацию оценивали по статистическим критериям отношения шансов (ОШ) с 95% доверительным интервалом (ДИ). Для проведения мета-анализа использовали программное обеспечение RevMan (Cochrane Collaboration, 5.3. Копенгаген). Для оценки связи четырех полиморфизмов rs2107425, rs2839698, rs217727, rs3741219 с риском развития рака молочной железы в мета-анализ были включены 9 исследований случай-контроль с общей выборкой из 6572 пациентов с раком молочной железы и 6968 доноров в контрольной группе. В результате исследования мы наблюдали связь между rs2839698 H19 и риском развития рака молочной железы в аллельной модели (ОШ = 1,33, 95% ДИ: 1,00- 1,76, Pz = 0,005, Pi2 = <0.00001). Кроме того, в рецессивной модели риск развития рака молочной железы также был ассоциирован с мутантным аллелем rs2839698 (ОШ = 1,33, 95% ДИ: 1,03-1,71, Pz = 0,03, Pi2 = 0,05). Значимых ассоциаций с риском развития рака молочной железы полиморфизмов rs2107425, rs217727, rs3741219 не обнаружили. Настоящий мета анализ показал, что полиморфизм rs2839698 H19 ассоциирован с риском развития рака молочной железы. The meta-analysis was aimed to evaluate the association between polymorphisms rs2107425, rs2839698, rs217727, rs3741219 of the H19 gene and the risk of developing of breast cancer. Publications were searched in the Google Scholar and PubMed databases until August 2023. The association was assessed by statistical odds ratio (OR) criteria with a 95% confidence interval (CI). RevMan software (Cochrane Collaboration, 5.3. Copenhagen) was used for the meta-analysis. To assess the association of polymorphisms rs2107425, rs2839698, rs217727, rs3741219 with the risk of developing breast cancer, 9 case-control studies with a total sample of 6572 patients with breast cancer and 6968 donors in the control group were included in the meta-analysis. As a result of the study, we observed an association between rs2839698 H19 and the risk of developing breast cancer when calculating the allelic model (OR = 1.33, 95% CI: 1.00-1.76, Pz = 0.005, Pi2 = <0.00001). In addition, analysis of the recessive model also showed that the risk of developing breast cancer was significantly associated in individuals with a mutation in the polymorphic allele rs2839698 (OR = 1.33, 95% CI: 1.03-1.71, Pz = 0.03, Pi2 = 0.05). When calculating genetic models for the rs2107425, rs217727, rs3741219 polymorphisms, no statistically significant associations with the risk of developing breast cancer were found. The present meta-analysis showed that the H19 rs2839698 polymorphism associated with the risk of breast cancer.

Over the past decade, the use of adoptive immunotherapy has emerged as a promising approach to treatment of various malignancies, including breast cancer. Cytokines are involved in the development, differentiation and homeostasis of the T cells. Despite the similarities in signaling pathways, the γc cytokines shape the T cell responses differently: IL-2 plays a major role in the development and maintenance of the regulatory T cells, while IL15 stimulates the proliferation and cytotoxic functions of the CD8+ T cells and NK- cells that leads to an enhanced antitumor response. It is of interest to study the combined effect of these cytokines in an experiment.

Background. Alkylresorcinols (ARs) are polyphenolic compounds of microbial origin with a wide spectrum of biological activities and are potentially involved in host immune functioning. The present study is aimed at evaluating alterations in AR content in blood serum and faeces from healthy donors and patients with lung cancer in connection with response to immune checkpoint inhibitor (ICI) therapy to estimate the regulatory potential of AR. Methods. Quantitative analysis of AR levels, as well as other microbial metabolites in blood serum and faeces, was performed using gas chromatography with mass spectrometric detection; estimation of lymphocyte subsets was performed by flow cytometry; faecal microbiota transplantation (FMT) from lung cancer patients after ICI therapy to germ-free mice was performed to explore whether the intestinal microbiota could produce AR molecules. Results. AR concentrations in both faeces and serum differ dramatically between healthy and lung cancer donors. The significant increase in AR concentrations in mouse faeces after FMT points to the microbial origin of ARs. For several ARs, there were strong positive and negative correlations in both faeces and serum with immune cells and these interrelationships differed between the therapy-responsive and nonresponsive groups. Conclusions. The content of ARs may influence the response to ICI therapy in lung cancer patients. ARs may be considered regulatory molecules that determine the functioning of antitumor immunity.

Scientists agree that three-dimensional tumor cell cultures reflect the in vitro biological features of malignant tumors to a greater extent than it is the case with the conventional monolayer cultures. In particular, the results of assessing the effectiveness of anticancer drugs obtained in three-dimensional cell models are considered as more accurate and consistent with the tumor response in vivo, where sensitivity is usually lower than that found in the in vitro experiments. Some of the most developed cellular models are structures obtained using three-dimensional bioprinting, which include not only malignant cells, but also biogels to imitate the chemical and mechanical properties of the extracellular matrix of the tumor.

Lung cancer is referred to as the second most common cancer worldwide and is mainly associated with complex diagnostics and the absence of personalized therapy. Metabolomics may provide significant insights into the improvement of lung cancer diagnostics through identification of the specific biomarkers or biomarker panels that characterize the pathological state of the patient. We performed targeted metabolomic profiling of plasma samples from individuals with non-small cell lung cancer (NSLC, n = 100) and individuals without any cancer or chronic pathologies (n = 100) to identify the relationship between plasma endogenous metabolites and NSLC by means of modern comprehensive bioinformatics tools, including univariate analysis, multivariate analysis, partial correlation network analysis and machine learning. Through the comparison of metabolomic profiles of patients with NSCLC and noncancer individuals, we identified significant alterations in the concentration levels of metabolites mainly related to tryptophan metabolism, the TCA cycle, the urea cycle and lipid metabolism. Additionally, partial correlation network analysis revealed new ratios of the metabolites that significantly distinguished the considered groups of participants. Using the identified significantly altered metabolites and their ratios, we developed a machine learning classification model with an ROC AUC value equal to 0.96. The developed machine learning lung cancer model may serve as a prototype of the approach for the in-time diagnostics of lung cancer that in the future may be introduced in routine clinical use. Overall, we have demonstrated that the combination of metabolomics and up-to-date bioinformatics can be used as a potential tool for proper diagnostics of patients with NSCLC.

Purpose of the study. Testing the protocol of obtaining cell spheroids of breast cancer cell cultures for bioprinting by growing in alginate drops.Materials and methods. Cells of breast cancer cell lines BT-20 and MDA-MB-453 were cultured in DMEM medium supplemented with 10 % FBS. Next, the cells were removed from the plastic using a trypsin-V ersene solution and resuspended in a sterile 2 % alginate solution in DPBS to the concentration of 105 cells/ml. Then the alginate solution with the cells was slowly dripped through a 30G needle into a sterile cooled solution of calcium chloride (100 mM) from a height of 10 cm. After polymerization, alginate drops were washed in DMEM and cultured for two weeks in DMEM with the addition of 10 % FBS at 37 °C and 5.0 % CO2.The spheroids formed in the alginate were photographed on the 3rd, 7th, 10th, and 14th days of cultivation, after which they were removed from the alginate by keeping in 55 mM sodium citrate solution with the addition of 20mM ethylenediaminetetraacetic acid (EDTA) and embedded in paraffin blocks according to the standard method, followed by histological examination.Results. Cellular spheroids were formed in both cell cultures already on the 3rd day of cultivation. From the 3rd to the 10th day in both cultures, a uniform growth of cell spheroids was observed with a gradual slowdown in the increase in the size of spheroids by the 14th day of cultivation. On the 10th day the proportion of cells that formed clones (more than 500 μm2 in size) was 25.2 % ± 7.1 % (n = 25) in the BT-20 culture and 38.5 % ± 9.9 % (n = 25) in MDA-MB-453 culture. On the 14th day, BT-20 culture was characterized by spheroids varying little in size and shape, with an average area of 1652 ± 175 µm2, having a dense structure with smooth edges. The spheroids in MDA-MB-453 culture turned out to be more loose and easily deformed, their size and shape varied noticeably, the average area of the spheroids was 2785 ± 345 µm2.Conclusion. The production of spheroids in alginate drops is inferior in speed to the methods of forming cell conglomerates in hanging drops or on microwells, but it surpasses these methods in productivity, which is comparable to the production of spheroids by constant medium stirring on low-adhesive substrates. In addition, the clonal nature of the obtained spheroids leads to an increase in research costs and thus limits their scalability.

3D печать - это новый многообещающий метод создания трехмерных клеточных конструкций для всевозможных биомедицинских исследований. К преимуществам использования 3D-биопечати в биомедицинской сфере относятся разработка персонализированного дизайна для каждого пациента, высокая точность и создание сложных структур. Основным компонентом 3D-биопечати являются биочернила, обеспечивающие биосовместимость, механическую стабильность, и высокое разрешение вовремя и после печати. Цель. Изучить влияние способа отверждения биочернил на основе метакрилата желатина и альгината на микроструктуру получаемого 3D конструкта и на морфологию инкапсулированных в него клеток рака молочной жедезы (РМЖ) ВТ20. В нашем исследовании мы использовали метод экструзионной 3D печати на биопринтере BIO X (Cellink, США) с биочернилами на основе альгината и метакрилата желатина GelMa, смешанные с клетками линии рака молочной железы BT-20 в соотношении 2:1. Напечатанные конструкты полимеризировали двумя способами химическим или фото- отверждением. После отверждения конструкты с клетками помещали в питательную среду DMEM с добавлением 10% FBS и культивировали при 37°С и 5,5% СО2. Затем образцы наблюдали и визуализировали с помощью микроскопа (Ti-S, Nikon, Япония). По истечении одной и двух недель культивирования часть конструктов фиксировали и заключали в парафиновые блоки. Далее по стандартной методике подготавливали срезы и окрашивали препараты гематоксилином и эозином. В результате мы спроектировали квадратные, 3-х слойные, конструкты с инкапсулированными клетками рака молочной железы. При создании 3D моделей опухолевого роста РМЖ с использованием биочернил на основе метакрилата желатина и альгината на наш взгляд, предпочтительным является фотоотверждение, так как оно позволяет создать губчатую микроструктуру из сообщающихся пор. Такая структура поддерживает миграцию клеток и способствует сохранению клеточной морфологии, приближенной к наблюдаемой in vivo.

National Medical Research Centre for Oncology, Ministry of Health of Russia, Rostov-on-Don, Russia Breast cancer (BC) is among most cancers worldwide. To date, research is being actively conducted to study substances that can influence on the phenotype of the tumor cells. In chemotherapy with various drugs, the tumor cells have an ATP deficiency that contributes to a decrease in their proliferative activity. In the course of numerous pharmacological studies, the natural compound berberine isolated from the roots of plants of the genus Berberis sp. [1] has engaged the researchers’ attention as a candidate for this purpose.

e15079 Background: Initially, berberine was used as an antimicrobial agent in traditional medicine, but its anticancer properties were later discovered. In addition to its cytotoxic effect, berberine also has the ability to inhibit cell motility, which has been demonstrated in some permanent cancer cell lines. The objective of this study was to assess berberine anti-migratory activity in permanent cell cultures compared to primary cell cultures which are generally thought to better reflect tumor characteristics. Methods: H1299 lung cancer, PC3 prostate cancer, and T98G glioma cells, as well as primary cell cultures of the corresponding cancer obtained in our laboratory, were planted in an amount of 15*104 cells per well of a 24-well plate (Biofil, China) in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA). After cell adhesion berberine was added in concentration 5 µM, and a wound healing assay was performed according to the standard procedure. Cell plates were continuously incubated and photographed in Lionheart FX imager (BioTek, USA) at 37°C and 5.0% CO2. The extent of cell migration was measured as the percentage reduction in wound area after 48 hours of incubation relative to baseline. Data are presented as Mean ± 95% confidence interval (n = 12). Results: The use of berberine at a concentration of 5 µM led to a significant decrease in cell motility in permanent cultures of lung cancer, glioma and prostate cancer. Namely, the reduction in the wound area after 48 hours of incubation with bereberine was 74.52±12.3% (compared with 94.56±6.2% in the control) in H1299 cell culture, 38.22±10.6% (compared with 83.89±15.5% in the control) in T98G cell culture, and 48.6±7.5% (compared with 69.56±8.1% in the control) in PC3 cell culture. The resulting difference between the control and experimental groups in permanent cell cultures was statistically significant at a significance level of 5% (df = 22). At the same time, the values of wound area reduction for primary cultures of the same cancers did not differ significantly in the control and under the influence of 5 μM berberine at the accepted level of significance. Namely, the reduction in the wound area after 48 hours of incubation with bereberine was 84.79±11.2% (compared with 81.47±15.3% in the control) in lung cancer primary cell culture, 94.64±5.1% (compared with 91.73±6.8% in the control) in glioma primary culture, and 62.63±5.8% (compared with 61.1±8.9% in the control) in prostate cancer primary cell culture. Conclusions: Permanent cell lines are more sensitive to berberine anti-migratory activity than primary cancer cell lines of the same localization.

We conducted this meta-analysis to estimate associations between CDKN2B antisense (CDKN2B-AS) rs2383207 polymorphism and susceptibility to atherosclerosis. A systematic literature research of Google Scholar and PubMed was performed to identify eligible studies. Overall, eight studies were included for meta-analyses. The association was assessed by statistical odds’ ratio (OR) with 95% confidence interval (CI). RevMan software (Cochrane Collaboration, 5.3. Copenhagen) was used for the meta-analysis. Pooled overall analyses showed that rs2383207 polymorphism was associated with the risk of atherosclerosis in the whole population. Additional analyses by ethnicity revealed that rs2383207 polymorphism was associated with susceptibility to atherosclerosis in Asians and Caucasians. Our results suggest that rs2383207, might serve as genetic biomarkers of atherosclerosis. Further, studies will be required to confirm the observed association.

The Biobank of the National Medical Research Center of Oncology is a multi-layered infrastructure with large collections of biological samples, complemented by extensive and well-annotated clinical and pathological patient data, including medical images, pathological histology, and molecular analysis of biosamples. To date, the biobank of the National Medical Research Center of Oncology contains collections of primary and immortalized cancer cell lines of human origin. The collection of primary cell lines was formed from samples of postoperative material taken during the removal of tumors of various localizations (breast cancer, prostate cancer, lung cancer). All cell lines underwent internal quality control for contaminants (exogenous viruses, mycoplasmas and bacterial L-forms), viability and were cultivated without antibiotics. On the basis of the collected samples, a significant number of projects in the field of biomedicine were carried out, the results of which are described in this article.

e15078 Background: It is known that berberine inhibits oxidative phosphorylation by suppressing respiratory complex I function, but the exact mechanism of this phenomenon has not yet been described. The aim of the study was to evaluate the effect of residues in positions 8 and 13 of berberine molecule on the energy metabolism of HeLa cells. Methods: HeLa cells were seeded in an amount of 2*104 cells per well in a Seahorse XFp Analyzer plate (Agilent, USA) in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA). After cell adhesion, the medium was decanted, and the medium with test substances at a concentration of 5 µM was added. Four variants of the experiment, 3 repetitions each, were set up: 1) control; 2) berberine without modification; 3) modification 1 (M1) with the addition of 7-nitrobenzo[c][1,2,5]oxadiazol-4-yl residue at position 13 and malononitrile residue to position 8; 4) modification 2 (M2) with the addition of dicyanovinyl residue at position 13. After 24 hours the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured on Seahorse XFp Analyzer (Agilent, USA) using a Seahorse XFp Cell Energy Phenotype Test (Agilent, USA), with 2 µM FCCP and 1 µM oligomycin. Results: Incubation with the original berberine resulted in a significant decrease in baseline OCR by 39.3±2.8% (M±SD) compared to control (α = 0.05, df = 4), but the maximum OCR did not change significantly. At the same time, a significant increase in the baseline level of glycolysis was observed proportional to the decrease in the baseline OCR, as can be seen from the increase in ECAR by 32.9±5.3% compared with the control (α = 0.05, df = 4). Berberine M1 modification led to a significant increase in baseline OCR by 49.1±2.1% compared with the control (α = 0.05, df = 4), which was also accompanied by a significant increase in maximum OCR by 22.8±3.8% (α = 0.05, df = 4), while the increase in the baseline level of ECAR by 9.7±4.2% was not significant. The addition of the M2 modification did not significantly change the measured parameters compared to the control. Conclusions: The introduction of a dicyanovinyl residue into the berberine molecule at position 13 abrogated its ability to inhibit cellular respiration, possibly due to disturbing its interaction with the proteins of the respiratory complex I. Introduction of the bicyclic residue 7-nitrobenzo[c][1,2,5]oxadiazol-4- yl at position 13 and a malononitrile residue at position 8 caused an increase in baseline and maximum respiration levels, that means thereby berberine activity reversal. This effect can be associated either with the ability to uncouple oxidative phosphorylation (similar to FCCP) or to enhance the expression of electron transport chain proteins.