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Purpose of the study. An analysis of changes in the expression of the VEGF neoangiogenic factor in the tumor tissue of patients with squamous cell carcinoma of the oral mucosa receiving targeted therapy with cetuximab and chemotherapy.Patients and methods. We performed an immunohistochemical study of tumor samples obtained from 60 patients with squamous cell carcinoma of the oral mucosa T3-4N0-1M0. The main group comprised 30 patients who received therapy with cisplatin and fluoruracil plus cetuximab. The control group included 30 patients receiving standard chemotherapy without targeted therapy. Each group was divided into two subgroups with different treatment efficacy: patients sensitive to treatment (n = 17 in the group with cetuximab and n = 12 in the group without cetuximab) and resistant to treatment (n = 13 in the group with targeted therapy and n = 18 in the group with standard chemotherapy).Results. Quantification of the VEGF expression demonstrated minimal numbers of vessels stained positively for this marker in the field of view in patients of the main group sensitive to chemotherapy and cetuximab. The value was 5.3 times lower than initial values, and 4.3 times lower than in the subgroup of patients resistant to the treatment (the data were statistically significant, р = 0.0132 and р = 0.0455, respectively). In the control group, patients who were sensitive to the treatment showed 1.4 times lower values than initially (р = 0.921), and patients who were resistant to the treatment had 1.1 times lower values than initial values (р = 0.936). The data were not statistically significant.Conclusions. The study showed that the number of microvessels in patients resistant to chemotherapy and cetuximab was 4.3 times higher than in patients with effective targeted therapy (р = 0.0455). The differences in the control group were not statistically significant.

Purpose of the study. An analysis of parameters of the blood coagulation system in breast cancer patients after coronavirus disease.Materials and methods. 50 breast cancer patients were divided into groups: the main group included 30 patients after coronavirus disease, the control group 1–20 patients without confirmed COVID‑19, and control group 2–20 non-cancer women after corona‑ virus disease. All cancer patients received appropriate chemotherapy. The following parameters were studied: activated partial thromboplastin time (APTT), prothrombin time (PT), international normalized ratio (INR), prothrombin index (PTI), fibrinogen, soluble fibrin monomer complexes (SFMCs), thrombin time (TT), antithrombin III, D-dimer and plasminogen, fibrin degradation products. Blood tests were performed 4–6 weeks after the infection and two negative PCR test results for COVID‑19.Results. Patients of the main group demonstrated differences in INR values after treatment in the subgroups with asymptomatic disease (Me = 1.24) and with mild symptoms (Me = 0.97) U = 10; Z = 2.766; р = 0.0057, in subgroups with asymptomatic disease (Me = 1.24) and with moderate to severe symptoms (Me = 0.98) U = 26.5; Z = 2.199; р = 0.027, and in TT values in subgroups with asymptomatic disease (Me = 14.5) and with moderate to severe symptoms (Me = 16.5) U = 18.5; Z = –2.725; р = 0.0064. The comparison of the parameters in patients after COVID‑19 before (Me = 0.83) and after treatment (Me = 0.4) showed differences in the D-dimer values in patients with moderate to severe disease U = 6.5; Z = –2.2861; р = 0.022 towards their decrease after the therapy. Differences were found in APTT values between the main group (Me = 30.65) and control group 1 (Me = 27.85) U = 119; Z = 3.574; р = 0.00035, in antithrombin values between the main group (Me = 94) and control group 1 (Me = 106) U = 112; Z = 3.713; р = 0.00021, and in SFMCs values between the main group (Me = 17) and control group 1 (Me = 8) U = 180.5; Z = 2.356; р = 0.018.Conclusions. Determination of plasminogen levels can become an independent factor in detecting thrombotic risk in cancer patients who recovered from COVID‑19. Previous infection with COVID‑19 should be considered an additional risk factor for venous thromboembolic complications for cancer patients.

Introduction. Erythropoietin (EPO) application is a pathogenetic method for anemia correction in cancer patients.The purpose of study. Clinical evaluation of the efficacy and safety of Eralfon® (epoetin alpha) in treatment for anemia in patients with malignant solid tumors during medical anticancer therapy.Materials and methods. We analyzed the data on anemia treatment with Eralfon® in 184 patients with malignant solid tumors receiving various medical anticancer therapies. Eralfon® was injected subcutaneously 12 000 IU 3 times per week or 40 000 IU once a week. Clinical antianemic effect, the time to maximum antianemic effect, adverse events (AE) were analyzed.Results. Patients were stratified by the grade of anemia, stages of treatment, presence of bone metastases, bleeding, previous medical and radiation anticancer therapies, dosage of Eralfon®. The time to effect was shorter in patients under 65. There were no significant differences in efficacy depending on the dosing regimen of Eralfon®. Efficacy was lower in patients with advanced tumors, especially in bone metastases. A history of tumor bleeding, chemotherapy and/or radiation therapy prolonged the period of hemoglobin recovery to normal values. Arterial hypertension and venous thrombosis were the most common AE associated with Eralfon®. Eralfon® 12 000 IU 3 times per week caused less frequent complications, with no cases of ossealgia and myalgia.Conclusion. Eralfon® demonstrated clinical efficacy in treatment for anemia in patients with solid malignant tumors receiving medical anticancer therapy. Dosage of 12 000 IU 3 times per week provided better control of the antianemic effect and adverse events.

Differential fluorescent staining is an effective tool widely adopted for the visualization, segmentation and quantification of cells and cellular substructures as a part of standard microscopic imaging protocols. Incompatibility of staining agents with viable cells represents major and often inevitable limitations to its applicability in live experiments, requiring extraction of samples at different stages of experiment increasing laboratory costs. Accordingly, development of computerized image analysis methodology capable of segmentation and quantification of cells and cellular substructures from plain monochromatic images obtained by light microscopy without help of any physical markup techniques is of considerable interest. The enclosed set contains human colon adenocarcinoma Caco-2 cells microscopic images obtained under various imaging conditions with different viable vs non-viable cells fractions. Each field of view is provided in a three-fold representation, including phase-contrast microscopy and two differential fluorescent microscopy images with specific markup of viable and non-viable cells, respectively, produced using two different staining schemes, representing a prominent test bed for the validation of image analysis methods.

IntroductionComplex gait disturbances represent one of the prominent manifestations of various neurophysiological conditions, including widespread neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. Therefore, instrumental measurement techniques and automatic computerized analysis appears essential for the differential diagnostics, as well as for the assessment of treatment effectiveness from experimental animal models to clinical settings.MethodsHere we present a marker-free instrumental approach to the analysis of gait disturbances in animal models. Our approach is based on the analysis of video recordings obtained with a camera placed underneath an open field arena with transparent floor using the DeeperCut algorithm capable of online tracking of individual animal body parts, such as the snout, the paws and the tail. The extracted trajectories of animal body parts are next analyzed using an original computerized methodology that relies upon a generalized scalable model based on fractional Brownian motion with parameters identified by detrended partial cross-correlation analysis.ResultsWe have shown that in a mouse model representative movement patterns are characterized by two asymptotic regimes characterized by integrated 1/f noise at small scales and nearly random displacements at large scales separated by a single crossover. More detailed analysis of gait disturbances revealed that the detrended cross-correlations between the movements of the snout, paws and tail relative to the animal body midpoint exhibit statistically significant discrepancies in the Alzheimer's disease mouse model compared to the control group at scales around the location of the crossover.DiscussionWe expect that the proposed approach, due to its universality, robustness and clear physical interpretation, is a promising direction for the design of applied analysis tools for the diagnostics of various gait disturbances and behavioral aspects in animal models. We further believe that the suggested mathematical models could be relevant as a complementary tool in clinical diagnostics of various neurophysiological conditions associated with movement disorders.

Abstract The emergence of new genes and functions is of paramount importance in the emergence of new animal species. For example, the insertion of the mobile element Tigger 2 into the sequence of the functional gene POU2F1 in primates led to the formation of a new chimeric primate-specific isoform POU2F1Z, the translation of which is activated under cellular stress. Its mRNA was found in all species of monkeys, starting with macaques. Analysis of the fragments of the Tigger2 copy corresponding to the human exon Z showed that the splicing sites of exon Z are homologous in humans and in most monkeys, with the exception of lemurs and galagos. The stop codon introduced into the mRNA by the Tigger2 sequence is present in all primates, starting with macaques. The internal ATG codon is also present in all primates, with the exception of lemurs and galagos. In the course of evolution, other MGEs, mainly of the SINE type, were inserted into the Tigger2 copy. In the course of evolution, both the location and the number of mobile SINE elements within the POU2F1 gene changed. Starting with macaques, the pattern of the arrangement of SINE elements within the Tigger2 copy in the studied region of the POU2F1 gene was fixed and then remained unchanged in other primates and humans, which may indicate its functional significance.

Background . Breast cancer is one of the most common malignancies and one of the leading causes of cancer deaths among women in the Russian Federation. The prognosis of the disease is largely determined by the biological subtype of the tumor. Her2-positive breast cancer is characterized by an aggressive course and unfavorable prognosis. The use of pertuzumab, a recombinant humanized monoclonal antibody to the human epidermal growth factor receptor 2, significantly improved immediate and long-term treatment outcomes. The aim of the study was to evaluate treatment outcomes in patients with Her2-positive molecular subtype of breast cancer receiving chemotherapy combined with a dual anti-Her2 blockade, and to assess the intensity and incidence of side effects. Material and methods . Between 2015 and 2018, 37 patients with Her2-positive breast cancer received chemotherapy combined with a dual anti-HER2 blockade (docetaxel 75 mg/m2 i/v on day 1 + trastuzumab 6 mg/kg (loading dose 8 mg/kg) i/v on day 1 + pertuzumab 420 mg (loading dose 840 mg) i/v on day 1 once every 3 weeks). The mean age of the patients was 45.6 ± 11.6 years. Results. Neoadjuvant pertuzumab in combination with trastuzumab and docetaxel resulted in pathological complete response in 12 % of patients and pathological partial response in 36 % of patients. Among patients who received the above neoadjuvant therapy regimen, disease progression was observed in 4 % of patients. The most common side effects were weakness and fatigue (5.4 % of cases) and grade I–II leukopenia (5.4 %). Conclusion. The study demonstrated the efficacy of antitumor therapy with pertuzumab in combination with trastuzumab and docetaxel as well as the absence of severe side effects associated with this treatment regimen in patients with Her2-positive breast cancer.

The POU2F1 gene, which plays an important role in regulating the mammalian genome and development, has both a ubiquitous (U) and a tissue-specific (L) promoter and is subject to intricate regulation. Regions of POU2F1 gene were found to contain multiple binding sites for its product POU2F1 (Oct-1), a transcription factor. Interspecies homology in these regions was found to exceed 90% among the human, mouse, rat, pig, and dog genomes, almost all of the Oct-1 binding sites being identical. Some of the sites cluster in the vicinity of each of the two alternative promoters, while others are in the 5' noncoding region 6 kb upstream of the transcription start site. The presence of Oct-1 at the sites was demonstrated by chromatin immunoprecipitation and the electrophoretic mobility shift assay (EMSA). A POU2F1 knockdown activated the U promoter and downregulated the L promoter in Namalwa cells, while Oct-1 overexpression exerted an opposite effect. Thus, Oct-1 acts via negative feedback to autoregulate the U promoter through low-affinity Oct-1 binding sites and positive feedback to autoregulate the L promoter through high-affinity canonical (oct) sites when increasing in concentration in a natural context.

Introduction:Squamous cell carcinoma of the oral cavity is one of the most common head and neck cancers with an aggressive course and high mortality rates.The aimof the study was to determine the EGFR expression levels in tumor tissues in patients with squamous cell carcinoma of the tongue and oral mucosa depending on the efficacy of the therapy.Material and methods:The study included 60 patients with squamous cell carcinoma of the tongue and oral mucosa T3-4N0-1M0. The main group included 30 patients receiving chemotherapy (cisplatin/fluorouracil) in combination with targeted therapy with cetuximab. The control group included 30 patients receiving chemotherapy without cetuximab. Both groups were divided into two subgroups: sensitive and resistant.Results:In treatment-resistant patients of the main group with cetuximab, the average EGFR expression was twice lower than the initial levels (p = 0.0080) and 1.7 times higher than in treatment-resistant patients of the control group (p = 0.0157). In treatment-sensitive patients, the average EGFR expression was 19.8 times lower (p = 0.0020) than initial values and 14.9 times higher (p = 0.0067) than in treatment-sensitive controls.Conclusions:A natural decrease in the EGFR expression in tumor tissues due to the targeted therapy was revealed. However, some patients were resistant to cetuximab, which dictates the need to search for predictors of targeted therapy efficacy in patients with locally advanced squamous cell carcinoma of the tongue and oral mucosa.

Aim. To study the prevalence of carriage of polymorphic allele variants of genes of blood coagulation factors in oncological patients. Materials and Methods. 213 Patients with morphologically confirmed oncological diseases were examined. Samples of genomic DNA of peripheral blood of the patients were examined. Using polymerase chain reaction (PCR), polymorphic sites of genes of hemostatic system were studied in real time: F2 (G20210А, rs1799963), F5 (G1691A, rs6025), F7 (G10976A, rs6046), F13 (G226A, rs5985), FGB G(-455)A (rs1800790), ITGA2-2 (C807T, rs1126643), ITGB3-b (Т1565С, rs5918), PAI-1 4G(-675)5G, rs1799889). Results. The prevalence of carriage of alternative allele of F2 (G20210А) polymorphic locus in the studied group was 1.6%, of F5 (G1691A) 3.5%, of F7 (G10976A) 13.4%, of F13 (G226A) 28.2%, of FGB G(-455)A 24.9%, of ITGA2-2 (C807T) 41.5%, of ITGB3-b (Т1565С) 15.5%, of PAI-1 4G(-675)5G 56.6%. A statistically significant increase in the frequency of risk alleles of F5 G1691A (р=0.0169), F13 G226A (р=0.0007), FGB G(-455)A (р0.0001) and ITGA2-2 C807T (р=0.0201) polymorphic loci was found in oncological patients as compared to the general population. In the same loci, except ITGA2-2 (C807T), statistically significant differences in the frequency of alternative alleles were found in different localizations of the oncological process. In 92.0% of patients, SNR combination was determined in different components of hemostatic system. Conclusion. Taking into account a high frequency of identification of risk alleles in all components of hemostatic system, it is reasonable to carry out additional research to determine the necessity of addition of antiaggregants to antithrombotic therapy in oncological patients.

Aim. Study of VEGF-A and TGF-β growth factors in tumor tissue bioptates in patients with squamous cell carcinoma of the tongue and floor of the oral cavity depending on the effectiveness of polychemotherapy targeted therapy with cetuximab. Patients and methods. The study included 30 patients with squamous cell carcinoma of the tongue and mouth floor mucosa (T3-4N0-1M0). All patients received 2 cycles of therapy: cisplatin 100 mg/m2 , intravenously, day 1, 5-fluorouracil 1000 mg/m2 /day, intravenously, 96-hour continuous infusion in combination with targeted therapy (cetuximab 400 mg/ m2 on day 1 in a loading dose, then 250 mg/m2 on days 8 and 15). Patients were divided into two subgroups: target sensitivity of patients (partial regression and stabilization) n = 17 and target resistance (progression) n = 13. Levels of growth factors VEGF-A and TGF-β were determined in tumor tissue bioptates by ELISA using standard test systems (Bender Med System, Austria). Statistical processing of results was performed using the Statistica 6.0 program (Stat-Soft, 2001). Results. Polychemotherapy with cetuximab in some patients (n = 13), antitumor therapy with cetuximab did not result in statistically significant changes in levels of VEGF-A, TGF-β and the VEGF-A/TGF-β ratio compared to the initial values. In other patients (n = 17), the studied markers in tumor tissue bioptates were statistically significantly different from the initial values: VEGF-A was decreased by 1.46 times, TGF-β by 2.96 times, while the VEGF-A/TGF-β ratio was twice elevated. Conclusions. The results on levels of growth factors VEGF-A and TGF-β, as well as the VEGF-A/TGF-β ratio, are of a certain prognostic value and can be used as criteria for evaluating the efficacy of antitumor therapy in patients with squamous cell carcinoma of the tongue and mouth floor mucosa.

The authors analysed the efficacy and safety of Nab-paclitaxel (Nab-R) monotherapy in patients with metastatic breast cancer with visceral crisis (VC) in the second- and further-line chemotherapy. The objective response rate (ORR) was 35.3% (6 of 17 persons). The most frequent side effects were general weakness, nausea, symptoms of peripheral neuropathy. The degree of toxicity did not exceed 1–2 in 60% of cases. The median time to progression was 7.8 months. (95% CI 6–10.6). The median overall survival for patients with VC was 14.9 months. (95% CI 12.0–16.9). Efficacy and controlled toxicity of Nab-P allows its use in pre-treated patients, including ones with VC.

The purpose of the study was to measure blood levels of insulin-like growth factors in patients with squamous cell carcinoma of the tongue and lower oral mucosa depending of the efficacy of cetuximab chemotherapy. The study included the data on 50 patients with squamous cell carcinoma of the tongue and lower oral mucosa T3-4N0-1M0. Patients received chemotherapy with cetuximab (n=30, main group) and without it (n=20, controls). Results were compared with the values in 20 non-cancer donors. Both groups of patients were divided into two subgroups: with stable disease and partial regression. Before treatment, blood serum levels of the two insulinlike growth factors IGF-1 and IGF-2 in patients were lower than in donors by 53.5% and 20.3%, respectively. A cycle of the standard chemotherapy in the subgroup with partial regression statistically significantly increased IGF-1 levels by 33% compared to the values before treatment, while IGF-2 was both within donor and initial values and the IGF-1/IGF- 2 ratio was similar to the initial level but 33% lower than in donors. Cetuximab chemotherapy in patients with partial regression normalized IGF-1 levels increasing it compared to initial values - by 87%. IGF-2 levels did not differ statistically significantly from their initial values and were 32.5% lower than in donors. The IGF-1/IGF-2 ratio was 58% higher than before treatment. Two therapy types demonstrated than cetuximab chemotherapy normalized IGF-1 in patients with partial tumor regression shown by the IGF-1 increase up to normal blood levels in effective treatment.

e13072 Background: Breast cancer (BC) is the most common cancer in women worldwide. The purpose of the study was to analyze immediate and long-term results of treatment in patients with locally advanced primarily inoperable HER2-negative breast cancer depending on the time of progression. Methods: The study included 104 women with locally advanced primarily inoperable HER2-negative breast cancer who developed an early disease progression within 6 to 12 months after standard combination treatment. The blood levels of circulating tumor cells (CTCs) were measured in all patients during the progression stage. Results: An analysis of the overall (OS) and event-free (EFS) survival showed that median EFS in patients with luminal B subtype was 9 months, in patients with triple-negative (TNBC) subtype 8 months. EFS at 6 months was 87.5% in luminal B subtype and 79.4% in TNBC, p = 0.37985. Median OS in patients with luminal B subtype was 25 months, in patients with TNBC 26 months. One-year OS was 100% in luminal B subtype and 93.9% in TNBC, p = 0.138. 2-year OS was 54.2% in luminal B subtype and 55.9% in TNBC, p = 0.697. 3-year OS was 37.5% in luminal B subtype and 41.2% in TNBC, p = 0.639. CTCs were detected in all patients, with higher levels in patients with luminal B subtype than in TNBC patients. HER+ CTCs were found in 30% of patients with luminal B subtype but not registered in patients with TNBC. Such results were obtained for the first time. Conclusions: The absence of significant differences demonstrated the aggressive disease course in patients with luminal BC with identified primary hormone resistance, similar to the TNBC course. The results should be taken into account when predicting the course of the disease and developing further treatment tactics.

e21509 Background: The limited range of patients clinically responding to checkpoint inhibitor therapy (CIT) is of interest and requires studying the immunological mechanisms underlying this phenomenon. It also remains to be decided or better clarified which biomarkers should be considered when selecting patients for immunotherapy. The purpose of this study was to evaluate some parameters of the cytokine profiles of peripheral blood in melanoma patients receiving CIT with the disease progression. Methods: The study included 11 patients aged 47-82 years with advanced melanoma. CIT (nivolumab 3 mg/kg once every 14 days or 240 mg once every 14 days) continued until the appearance of reliable signs of the disease progression (in combination with symptomatic deterioration of the patient's condition). The levels of cytokines (TNF-α, IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ) in the blood serum of patients were determined by ELISA before treatment, after 4 and after 8 injections. The results were expressed as specific content (units/mg). Results: All patients developed disease progression after immunotherapy. CIT was accompanied by different changes most pronounce after the 8th CIT cycle. Levels of IL-1β, IFN-γ, IL-10 and α-TNF exceeded the values before CIT by 118%, 131%, 84% and 20%, respectively (5.2 [4.37; 6.1] versus 2.0 [0.44;4], p≤0.041, 3.9 [3.7;4.5] vs. 1.7 [0.5;3.5], p≤0.021, 6.5 [5.8 ;8.2] vs. 3.5 [3.2;5.4], p≤0.04; 4.1 [3.9;4.9] vs. 3.4 [3.2;3.9] pg/ml, p≤0.047). Levels of IL-6, on the contrary, decreased by the 8th CIT cycle by 32%: 5.1 [3.5; 6.1] versus 7.5 [6.2; 22.3] pg/ml, p≤ 0.045. Levels of IL-2 and IL-8 did not change. Conclusions: The revealed significant increase in levels of IL-1β, IFN-γ, IL-10 and α-TNF, as well as the IL-6 decrease, may indicate certain features of the cytokine profile of melanoma patients during the treatment, which may affect the imbalance of regulatory molecules and contribute to the disease progression.

e18072 Background: Clinical course and aggressiveness of a tumor, as well as the subtle mechanisms of its pathogenesis are related to vascularization, which in turn is determined by the ability of transformed cells to synthesize a wide range of biologically active substances. Among modern approaches to cancer treatment, special attention is paid to anti-EGFR agents reducing tumor vascularization. The purpose of this study was to analyze VEGF expression in tumor vessels in patients with squamous cell carcinoma of the tongue and oral mucosa developing cetuximab resistance. Methods: The study included 45 patients with squamous cell carcinoma of the tongue and oral mucosa T3-4N0-1M0. The patients were divided into 3 groups: group 1 – all patients before treatment; group 2 – patients after standard chemotherapy with cisplatin and fluorouracil; group 3 – patients after chemotherapy in combination with targeted therapy with cetuximab. Immunohistochemical study was performed on sections from paraffin blocks of tumors using monoclonal murine anti-VEGF antibodies (clone VG1, Thermo Scientific) diluted 1:150. The Reveal Polyvalent HRP-DAB Detection System was used. The significance of differences was assessed using the Mann Whitney and Wilcoxon tests calculated using the STATISTICA 10.0 program (StatSoftInc., USA). Results: Changes in the density of vessels stained with the VEGF marker were studied in tumor cells of patients in all groups. Patients in group 1 had from 3 to 10 vessels in one field of view; in group 2 – from 2 to 6 vessels in 70% of cases (21) and up to 15 vessels in 30% of cases (9); in group 3 – only solitary vessels. The average numbers of microvessels stained with the VEGF marker per field of view were 6.5±1.52; 4.5±1.71; 1.0±0.28, respectively. Quantification of intratumoral microvascular density showed that the minimum number of microvasculature vessels in the field of view was observed in patients who received chemotherapy and targeted therapy with cetuximab. The value was 5 times lower compared to initial values, and 4 times lower than in patients who received only standard chemotherapy (the Mann Whitney test, p < 0.05). The statistical analysis according to the Wilcoxon test on the effect of targeted therapy (comparison between groups 2 and 3) revealed a statistically significant effect of cetuximab for this marker (p = 0.028). Conclusions: Immunohistochemical analysis of the VEGF neoangiogenesis factor in squamous cell carcinoma of the tongue and oral mucosa confirmed the effectiveness of targeted therapy with cetuximab.

e21068 Background: Lung cancer (LC) is one of the most commonly diagnosed cancers and leading causes of cancer death worldwide. A limited clinical benefit of checkpoint inhibitor therapy (CIT) remains an important issue. Various units of the immune system are involved in both antitumor immunity and the development of CIT-induced side effects. Cytokines play a special role in these phenomena. The purpose of this study was to evaluate the cytokine profiles of peripheral blood in patients with non-small cell lung cancer (NSCLC) receiving CIT. Methods: The study included 32 NSCLC patients aged 41-83 years. CIT (pembrolizumab 2 mg/kg once every 21 days or 200 mg once every 21 days) continued until the appearance of reliable signs of the disease progression (in combination with symptomatic deterioration of the patient's condition). The effect was assessed by RECIST 1.1. The levels of cytokines (TNF-α, IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ) in the blood serum of patients were determined by ELISA before treatment, after 4 and after 8 injections. The results were expressed as specific content (units/mg). Results: The disease progression was registered in 14 patients, complete response in 7, partial response in 3 and stabilization in 8 patients. Only TNF-α was increased after CIT by 39% (11.3 [10.5; 15.4] vs. 8.1 [7.37;10.2] pg/mL, p≤0.037), while levels of IL-2, IL-4, IL-6 and IL-10 were lower than the initial values by 96%, 100%, 36% and 56%, respectively (0.06 [0.03; 0.45] vs. 1.65 [1.2;2.3], p≤0.034; 0 [0; 0.02] vs. 0.09 [0.05; 1.1], p≤0.031; 14.5 [8.5; 15.9] vs. 22.9 [18.1; 24.3], p≤0.042; 3.3 [2.7; 4.1] vs. 7.5 [6.4; 8.3] pg/mL, p≤0.038). No changes were found in the levels of IL-1β and IL-8 in the peripheral blood. Conclusions: Immunotherapy with pembrolizumab in the studied cohort of NSCLC patients after the 8th CIT cycle was characterized by a decline in the levels of cytokines (IL-2, IL-4, IL-6 and IL-10) determining the development of the cellular and humoral immune response, which may contribute to the treatment results.

The POU2F1 gene, which plays an important role in regulating the mammalian genome and development, has both a ubiquitous (U) and a tissue-specific (L) promoter and is subject to intricate regulation. Regions of POU2F1 gene were found to contain multiple binding sites for its product POU2F1 (Oct-1), a transcription factor. Interspecies homology in these regions was found to exceed 90% among the human, mouse, rat, pig, and dog genomes, almost all of the Oct-1 binding sites being identical. Some of the sites cluster in the vicinity of each of the two alternative promoters, while others are in the 5' noncoding region 6 kb upstream of the transcription start site. The presence of Oct-1 at the sites was demonstrated by chromatin immunoprecipitation and the electrophoretic mobility shift assay (EMSA). A POU2F1 knockdown activated the U promoter and downregulated the L promoter in Namalwa cells, while Oct-1 overexpression exerted an opposite effect. Thus, Oct-1 acts via negative feedback to autoregulate the U promoter through low-affinity Oct-1 binding sites and positive feedback to autoregulate the L promoter through high-affinity canonical (oct) sites when increasing in concentration in a natural context.

The emergence of new primate-specific genes is an essential factor in human and primate brain development and functioning. POU2F1/Oct-1 is a transcription regulator in higher eukaryotes which is involved in the regulation of development, differentiation, stress response, and other processes. We have demonstrated that the Tigger2 transposon insertion into the POU2F1 gene which occurred in the primate lineage led to the formation of an additional exon (designated the Z-exon). Z-exon-containing primate-specific Oct-1Z transcript includes a short upstream ORF (uORF) located at its 5’-end and the main ORF encoding the Oct-1Z protein isoform (Pou2F1 isoform 3, P14859-3), which differs from other Oct-1 isoforms by its N-terminal peptide. The Oct-1Z-encoding transcript is expressed mainly in human brain cortex. Under normal conditions, the translation of the ORF coding for the Oct-1Z isoform is repressed by uORF. Under various stress conditions, uORF enables a strong increase in the translation of the Oct-1Z-encoding ORF. Increased Oct-1Z expression levels in differentiating human neuroblasts activate genes controlling stress response, neural cell differentiation, brain formation, and organogenesis. We have shown that the Oct-1Z isoform of the POU2F1/Oct-1 transcription factor is an example of a primate-specific genomic element contributing to brain development and cellular stress defense.

Thapsigargin, the SERCA ATPase inhibitor, effectively suppresses the expression of metastasis marker S100A4 in breast cancer cells MDA-MB231. It has been demonstrated that transcription of the S100A4 gene is controlled by Ca2+-signaling pathways. It has been shown that synthesis of S100A4 mRNA and protein in the MDA-MB231 cell line is effectively inhibited by thapsigargin at a concentration of 0.4-4 µM, while preserving cell survival. We assume that a change in gene transcription in response to the disruption of Ca2+ homeostasis plays a direct role in the remodeling of Ca2+-signaling pathways.

Thapsigargin, the SERCA ATPase inhibitor, effectively suppresses the expression of metastasis marker S100A4 in breast cancer cells MDA-MB231. It has been demonstrated that transcription of the S100A4 gene is controlled by Ca 2+ -signaling pathways. It has been shown that synthesis of S100A4 mRNA and protein in the MDA-MB231 cell line is effectively inhibited by thapsigargin at a concentration of 0.4-4 µM, while preserving cell survival. We assume that a change in gene transcription in response to the disruption of Ca 2+ homeostasis plays a direct role in the remodeling of Ca 2+ -signaling pathways.

Thapsigargin (SERCA ATPase inhibitor) inhibited the S100A4 metastatic marker expression in MDA-MB231 breast cancer cells. We found that S100A4 gene transcription is regulated by Ca

e18563 Background: Cetuximab is a monoclonal antibody directed against the EGFR receptor, able to block the EGFR signaling pathway and also indirectly affect the secretion of some growth factors. The search for biomarkers responsible for cetuximab resistance is an urgent task. The purpose of the study was to analyze VEGF-A and TGF-β levels in tumor tissue bioptates in patients with squamous cell carcinoma of the oral cavity receiving chemotherapy (CT) and cetuximab. Methods: The study included 30 patients with HNSCC St III-IV, T3-4N0-1M0. All patients received 2 cycles: cisplatin 100 mg/m2, intravenously, day 1, 5-fluorouracil 1000 mg/m2/day, intravenously, 96-hour continuous infusion, in combination with targeted therapy cetuximab 400 mg/m2 on day 1 in a loading dose, then 250 mg/m2 on days 8 and 15). Patients were divided into two groups: with response to cetuximab and CT (partial regression and stabilization) n = 17 and progression considered (resistance to the treatment) n = 13. Levels of VEGF-A and TGF-β were determined in tumor tissue bioptates by ELISA using standard test systems (Bender Med System, Austria). Statistical processing of results was performed using the Statistica 6.0 program (Stat-Soft, 2001). Results: CT and cetuximab in patients with resistance did not result in statistically significant changes in levels of VEGF-A, TGF-β and the VEGF-A/TGF-β ratio in tumor tissue bioptates, compared to the initial values. The studied markers in tumor tissue bioptates in patients with response to CT were statistically significantly different from the initial values: VEGF-A was decreased by 1.46 times, TGF-β by 2.96 times, while the VEGF-A/TGF-β ratio was twice elevated (p < 0.05). Conclusions: The results on levels of VEGF-A and TGF-β and the VEGF-A/TGF-β ratio are of a prognostic value and can be used for evaluating the efficacy of cetuximab and CT in patients with HNSCC.

e15514 Background: The standard treatment of squamous cell carcinoma of the tongue and the mucosa of the bottom of the oral cavity (SCCTOM) is chemotherapy (CT) in combination with cetuximab (mC, anti-EGFR antibody). However, not all patients manage to achieve positive results. A common cause of resistance to anti-EGFR antibodies is the constitutive activation of mediators of the underlying signaling pathways. Therefore, the aim of the study was to analyze mutations in the KRAS using the Digital Droplet PCR method in patients with SCCTOM on the background of CT in combination with targeted therapy or standard CT. Methods: The study used the QX200 system (Bio-Rad) and the ddPCRtm KRAS Screening Multiplex Kit (7 mutations). The study included 60 patients with T3-4N0-1M0 SCCTOM. The main group consisted of 30 patients (cisplatin, 5-fluorouracil+cetuximab). The control group consisted of 30 patients (CT). For research, cfDNA from blood plasma was used. Analysis of the data was performed using QuantaSoft v1.7.4. Results: The frequency of the KRAS mutant type (mtKRAS) is 43% before treatment and 32% after treatment (n = 60). In the group of patients CT+mC the incidence of mtKRAS before treatment was 40%, after treatment - 20%. In the group of patients CT without mC the incidence of mtKRAS before treatment was 47%, after treatment - 43%. Decrease of 20% (p = 0.00089) in the frequency of mtKRAS after CT+mC was found, while it was 23% (p = 0.00009) lower than the frequency of mtKRAS after CT without mC. After treatment, in the CT+mC group of patients, the number of mtKRAS DNA copies increased 1.5 times (p = 0.048). Analysis of clinical response to therapy allowed us to divide the main and control groups of patients into 2: sensitive and resistant to therapy. After treatment, a change in the mtKRAS ratio was not found in the subgroup sensitive to mC compared with the period before treatment, while the frequency of mtKRAS was reduced by 3 times (p = 0.009). In the mC-resistant subgroup, the mtKRAS ratio increased 1.9 times compared to the period before treatment (p = 0.009), while the frequency of mtKRAS increased 3.5 times compared with the mC-sensitive group (p = 0.0045). Conclusions: Prior to treatment, patients resistant to CT+mC were characterized by an increased occurrence of mutations in the KRAS and an apparently large number of tumor cells carrying mutations. The use of mCs caused a change in the direction of the clonal evolution of tumor cells - leading to an increase in the number of cells carrying mtKRAS (conditions were created for the elimination of cells with mutation in EGFR and KRAS wild type).

90 Background: Complete or partial deficiency of the DPD enzyme due to genetic polymorphisms of the DPYD gene causes acute toxicity of fluoropyrimidines, which are widely used in combination chemotherapy regimens for various malignant neoplasms. The purpose of the study: to identify polymorphisms of the DPYD gene significant for 5-fluorouracil-induced toxicity. Methods: Venous blood samples from Caucasian patients were used to identify alleles of *2А rs3918290, 5* rs1801159, *13 rs55886062 and rs67376798 DPYD by RT-PCR and direct sequencing. Inclusion criteria were: verified diagnosis of gastrointestinal tumors, age>18 years old, fluoropyrimidine-containing regimes of treatment. Results: 104 pts were included, 54% were female. Mean age-61 years. Colorectal cancer was found in 80.7% pts, non-colorectal in 19.3% pts. Hematological and non-hematological toxicity Gd.3-4 was found in 24% pts. Allele * 5rs18011595, which causes enzyme deficiency was found in 28% of patients, (frequency was 0.281) which is significantly higher than the population frequency of the allele charactering for Caucasoid population (p

Changes in the expression level of Oct-1A, Oct-1L, Oct-1X, and Oct-1Z isoforms and CD14 surface antigen during differentiation of HL-60 monocytic cells induced in vitro by dimethyl sulfoxide were studied, and the expression level of the four Oct-1 isoforms in vivo in human monocytes was determined.

It is demonstrated how to apply correlation and regression analysis for the purpose of evaluating economic activity of an enterprise. The basic regression models and examples of practical application thereof are provided.

e12589 Background: Better efficiency and safety of Nab-P is known to be due to the advantages of pharmacokinetics and pharmacodynamics of the drug. The fraction of unbound paclitaxel in plasma is 2.6-fold higher with Nab-P than with P. The purpose of the study was to assess efficacy and safety of Nab-P in ≥2nd lines of chemotherapy (CT) in patients (pts) with mBC with VC. Methods: Inclusion criteria: mBC with VC, ≥ 2 line of CT, HR+, HER2neu+ or triple negative type (TN), normal liver and renal function. Pts received Nab-P 260 mg/m2 q 21 days. Pts with Her2/Neu + received trastuzumab 6 mg/kg q 21 days. Results: 32 mBC pts, mean age 51.34 years (CI 35-76) were included. Among 32 pts Her2/Neu + had 5 (15.6%), TN – 9 (28.1%), HR+ 18(56.3%). 13(40.6%) pts received Nab-P as 2 line of CT, 13 (40.6%) pts, as 3-4 lines, 7(21.9%) as ≥5 lines. Pts were divided into 2 groups: group A - pts with VC 17(53.1%), group B - without VC 15(46.9%). Groups were comparable by age, number of previous CT lines, immunohistochemical types of BC. Group A had 2.47±0.32 CT lines (CV51.83%), including 8(47%) pts, who had received paclitaxel (P) in previous CT lines. Group B had on average 2±0.27 CT lines (CV53.03%). The objective response rate (ORR) was 37.5%(12 pts). In the group A ORR was 35.3% (6 pts). Out of 8 pts with VC, who had received P in previous lines, ORR was achieved in 3 pts (37.5%). In the group B ORR was 33.34%(5 pts). TTP was 6.8 mos (95% CI 6.1-13.7) for all pts, in the group A – 5.7 mos (95% CI 5.9-10.2), in the group B 6 mos (95% CI 6.5-13.7) (p = 0.046). Median 3-year OS of pts in group A was 18.9 mos (95% CI 4.3-22.4), in group B - 22.5 mos (95% CI 8.2-26.7) (p = 0.038). Toxicity did not differ significantly in pts in groups A and B and was manageable. Pts, who previously received P, had no new or severe toxicity. Conclusions: The data obtained confirms the possibility of Nab-P application in pts with VC. The use of Nab-P in pts treated with taxanes in previous lines is supposed for further study.

e17524 Background: Mutations in KRAS are known to be predictors of anti-EGFR therapy efficacy. Besides the mutation status of the treatment-naive tumors the changes in the level of mutant DNA during medical treatment are also of clinical interest for the individualization of the treatment. The purpose of the study was to detect somatic single nucleotide polymorphism (SNP) activating mutations in KRAS exon 2 in patients (pts) with OMTC undergoing chemotherapy (CT)±Cetuximab (Cet). Methods: Extracellular DNA samples were obtained from plasma of pts with OMTC (T3-4No-1Mo) undergoing CT (CDDP+5FU)+Cet (group A) before treatment and in 2 weeks after. Patients undergoing CT (CDDP+5FU, no Cet) were included in Group B. 7 activating mutations in KRAS exon 2 were detected by Droplet Digital PCR method (DD PCR) with "KRAS Screening Multiplex Kit" (BioRad, USA). Results: Samples from 28 pts were included. The frequency of SNP activating in KRAS exon 2 before treatment was 25%. In the group A (n = 12) KRAS mutations were not found or the level of mutant DNA did not exceed 1%. No significant changes were found in this group after treatment. The group B (n = 16) had some KRAS variations the majority of which did not change after treatment. However, 2 pts had a sharp increase in mutant DNA: from 0.03% before CT in both cases to 17.04% and 56.38% -after treatment (p < 0.01). Conclusions: Medical treatment, especially CT, could cause the changes in activating mutations in KRAS exon 2 in pts with OMTC. Revealing of these changes is of great importance for the following treatment options. Detection of mutant extracellular DNA by DD PCR could be valuable to monitor their status during the treatment.

e17523 Background: Treatment of advanced HNSCC (T3 and T4) involves the use of anti-EGFR monoclonal antibodies targeting the EGF-EGFR and VEGF-VEGFR cascades responsible for the cell proliferation, differentiation, invasion, metastasis, angiogenesis and apoptosis. The purpose of the study was the search for biomarkers of resistance to cetuximab in HNSCC patients. Methods: Tumor biopsy was performed in patients with HNSCC (n = 40) (Т3-4N0-1M0) before and after two cycles of chemotherapy with cetuximab - 400 mg/m2 administered on day 1 and 250 mg/m2 per week, cisplatin 100 mg/m2 on day 1, fluorouracil 1000 mg/m2 - 96-hour continuous infusion. Patients were divided into group 1 (progression, n = 11) and group 2 (stabilization and partial remission, n = 29). Levels of VEGF-A (pg/mL) (BenderMedSystem, Austria) and TGF-β (pg/mL) (eBioscience, USA) were determined in biopsy samples by ELISA, and the VEGF-A/TGF-β ratio was calculated. Statistical analysis of the results was performed using the Statistica 10.0 program with M±m determination. Differences were considered significant at p < 0.05. Results: Before treatment, levels of VEGF-A in biopsy samples were 634.8±37.2, TGF-β – 1092.5±71.9, and VEGF-A/TGF-β – 0.58±0.03. After 2 cycles of chemotherapy with cetuximab, results in group 1 for VEGF-A were 680.3±41.2, TGF-β – 1773.7±111.4, and VEGF-A/TGF-β – 0.384±0.021; in group 2: VEGF-A – 487.7±26.1, TGF-β – 572.3±34.7, and VEGF-A/TGF-β – 0.85±0.043. Thus, after 2 cycles of chemotherapy with cetuximab, TGF-β in group 1 increased by 1.6 times and VEGF-A/TGF-β – by 1.5 times (p < 0.05). In group 2, VEGF-A decreased by 1.3 times, TGF-β by 1.9 times, and VEGF-A/TGF-β increased by 1.46 times (p < 0.05). Conclusions: Decreased levels of markers and increased VEGF-A/TGF-β can predict cetuximab efficacy. Increased TGF-β and reduced VEGF-A/TGF-β allow predicting resistance to chemotherapy with cetuximab.

e12625Background: The data on TNF - drugs used in neoadjuvant treatment in breast cancer patients especially on its effect on survival is very few. Meanwhile some trials show that immunotherapy cou...

e18098Background: The impact of viral infections of tumor tissues on the effectiveness of antitumor treatment in patients with laryngeal cancer is poorly studied. Methods: Tumor tissues of 26 patie...

e14505Background: We monitored some biochemical parameters to assess hepatotoxicity of anthracycline chemotherapy in patients with breast cancer. Methods: The study included 30 patients (32-72 year...

e16211Background: The purpose of the study was to analyze the survival of patients with pancreatic cancer II-IV stages depending on the regimen used in the first line. Methods: The data of adults t...

e14506Background: The purpose of the study was to assess changes in biochemical markers of cardiotoxicity in anthracycline chemotherapy for breast cancer. Methods: The study included 30 pts (32-72 ...

Aim: Although breast cancer is a tumor sensitive to drug therapy, there is an urgent need for an effective treatment for pts with TNmBC. The study was performed to assess the efficacy and safety of CP combination in TNmBC. Methods: Elegibility criteria included measurable TNmBC, multiple metastases to visceral organs ≥ 2, ECOG ≤ 2, adequate liver, kidney and bone marrow function, no brain metastases. P was administered 175 mg/m2 on D1 and C AUC5 on D1 q3w up to 6 cycles. Treatment was continued until progression or unacceptable toxicity. Endpoints include response rate, OS, PFS and safety. Results: Between 2009-2013 81 pts with TNmBC were recruited. Mean age was 54.9 ± 3.4 yrs. All pts had multiple synchronous metastases to visceral organs ≥2 disease sites: lung 64,2% (52), lymph nodes 54.3% (44), bones 59.3% (48), liver 72.8% (59). 35.8% (29) pts received the 1st line and 64,2% (52) - the 2nd line chemotherapy (CT). RR was 80.2% (65) including 6.2% CR (5), 24.5% PR (20) and 49.4% SD (40). Most common toxicities were: nausea 1/2Gd-51.9%(42), vomiting 2Gd-13.6%(11), fatigue 1/2Gd-54,3%(44), bone pain 1/3Gd–69.1%(56), neutropenia 2/3Gd-32.1%(26). All kinds of toxicities were controlled by symptomatic medication if necessary and did not lead to CT delay. In the 1st CT line OS was 13.3 ± 2.4mos, PFS - 7.4 ± 1.5mos. In the CT 2st line OS was 8.2 ± 1.8mos, PFS - 4.6 ± 0.9mos. Conclusions: CP combination had a satisfactory efficiency and well-controlled toxicity. The favorable RFS and OS allow its use widely as clinical option in the 1-2 line of treatment of TNmBC pts with massive visceral metastases. Disclosure: All authors have declared no conflicts of interest.

e18564 Background: IGF-1 and IGF-2 are powerful mitogenic factors with anti-apoptotic effect. Their overproduction leads to neoangiogenesis and metastasis. Monitoring the level of these factors can serve as a potential predictive tool. The purpose of the study was to study blood levels of IGF-1 and IGF-2 in patients with squamous cell carcinoma (SCC) of the oral cavity depending of the efficacy of chemotherapy (CT) and cetuximab. Methods: The study included 50 patients with squamous cell carcinoma of the oral cavity T3-4N0-1M0, stage III-IV. The main group (n = 30) received 2 CT cycles with cetuximab: cisplatin 100 mg/m2, intravenously, day 1, 5-fluorouracil 1000 mg/m2/day, intravenously, 96-hour continuous infusion, in combination with cetuximab 400 mg/m2 on day 1 in a loading dose, then 250 mg/m2 on days 8 and 15. The control group (n = 20) received 2 CT cycles: intravenous cisplatin 100 mg/m2 on day 1, intravenous 5-fluorouracil 1000 mg/m2/day, 96-hour continuous infusion on days 1-4. Results were compared with the values in 20 non-cancer donors. Levels of IGF-1 (mcg/L) and IGF-2 (ng/mL) were measured in the blood serum by ELISA using standard test systems (Mediagnost, Germany). Statistical analysis of results was performed using the Statistica 6.0 program (Stat-Soft, 2001). Results: Before treatment, levels of IGF-1 and IGF-2 in patients were lower than in donors by 53.5% and 20.3%, respectively. In patients with partial regression after CT, IGF-1 levels significantly increased by 33% (p < 0,05), compared to the values before treatment, while IGF-2 was both within donor and initial values and the IGF-1/IGF-2 ratio was similar to the initial level but 33.3% lower than in donors. The patients in the main group with partial regression normalized IGF-1 levels increasing it compared to initial values – by 87%. IGF-2 levels did not differ statistically significantly from the initial values and were 32.5% lower than in donors. The IGF-1/IGF-2 ratio was 58% higher than before treatment. Conclusions: CT and cetuximab normalized blood levels of IGF-1 in oral cancer patients with partial tumor regression.