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Background. The progression of malignant tumors, including vulvar cancer, largely depends on the state of the extracellular matrix and immune microenvironment of the tumor, the study of which is an urgent problem of oncology to identify the most prognostically significant of them. Aim. To evaluate the expression of extracellular matrix and local immunity factors, as well as their relationships in vulvar cancer with different prevalence. Material and methods. A retrospective study on vulvar tumors in 56 patients with stage I (n=20), stage II (n=26) and relapses (n=10) was performed. The expression of markers of the extracellular matrix (matrix metalloproteinases MMP-2 and MMP-9, tissue inhibitor of metalloproteinase-2 TIMP-2) and local immunity (FOXP3, CD68, CD206) was determined by immunohistochemical method. Statistical processing was performed using the MannWhitney U-test and multiple correlation analysis with the calculation of paired and partial R coefficients. Results. The depth of invasion proved to be more informative for detecting multidirectional differences in the expression of MMP-2 and TIMP-2 than the stage. Infiltration by CD68+-macrophages in the stroma increased in parallel with the stage and did not differ at different depths of invasion; infiltration by macrophages M2 (CD206+) increased at stage II compared with stage I (Me 13 and 5, respectively, p=0.03). Differences were noted only in the stroma of tumors, as well as an increase in T-regs (FOXP3) with an increase in the depth of invasion (Me 25 and 8, respectively, p=0.0359). As the prevalence of the tumor developed, the number of correlation relationships between the parameters of local immunity of the parenchyma and tumor stroma increased: 2 statistically significant correlations that do not depend on covariates, 7 that depend on the stage, and 6 that depend on the depth of invasion, were found. Correlations of factors of the immune microenvironment with the extracellular matrix weakened. Conclusion. The levels of MMP-2, TIMP-2, CD68+, CD206+, FOXP3 reflect the progression of vulvar cancer and can be used to predict the course of the disease; infiltration of tumors by macrophages, T-regs characterizes the stage of the primary tumor rather than recurrence.

Background. Human papillomavirus (HPV) has been established to be the etiological factor of cervical cancer (CC). HPV infection and CC progression involve the direct participation of the E6 oncoprotein. Aim. An analysis of the E6 oncoprotein levels in tissues of the tumor and its perifocal area in HPV-associated cervical squamous cell cancer as an objective indicator of the effect of treatment depending on preoperative chemotherapy. Material and methods. The study included clinical and laboratory data of 237 patients with high-risk HPV infection of the cervix. The patients were divided into 4 groups: two main groups (CC T2а2–2bN0–1M0) and two control groups. Patients in the main group 1 (n=84) received standard neoadjuvant chemotherapy (NACT), in the main group 2 (n=93) — modified NACT with prior plasmapheresis session and a parallel course of nonspecific immunotherapy with Allokin-alpha. Control group 1 (n=40) included patients with CC T1b2–2a1N0–1M0, surgical treatment; control group 2 (n=20) — HPV-positive patients without CC. Levels of E6 were measured in samples of the cervical tumor and perifocal tissues. Results. The lowest levels of the E6 oncoprotein were registered in the group of HPV-positive patients without CC. After modified NACT, E6 levels in tumor tissues remained 4.6 times higher than in intact tissues, and even so, these patients demonstrated minimal E6 levels compared to other CC patients. E6 in tumor tissues was significantly lower than in main group 1 (by 3.3 times) and 8 times lower than in control group 1. E6 levels in the perifocal tissues of patients in main group 2 were 1.9 times lower than in the corresponding tissues of patients in main group 1 and 2.2 times lower than in control group 1. Conclusions. Inclusion of plasmapheresis and inducers of endogenous interferonogenesis into neoadjuvant treatment for cervical cancer can be considered pathogenetically justified, since it affects the key unit of cervical carcinogenesis.

Rationale: Ovarian cancer is the leading death cause in gynecological malignancies. More than 70% of the patients are diagnosed with progressing disease extending to outside the true pelvis. The 5-year survival of ovarian cancer patients remains low (about 47%) due to frequent relapses and drug resistance. Identification of markers for early diagnosis and relapse prediction could improve the outcomes of the disease.Aim: To assess relative copy number of cancer-associated genetic loci c-Myc, CDK12, CDKN1B, PRPF19, ERBB2, DDB1, GAB2, COL6A3 in the tumor cells of ovarian cancer, in order to identify potential prognostic oncomarkers in ovarian cancer patients.Materials and methods: The study included 50 women aged 27 to 70 years with ovarian cancer T1-3cN0-1M0-1, Gr. 2 (stages I—IV), who received their elective treatment in the National Medical Research Centre for Oncology in 2015 to 2019. The study was based on samples of genomic DNA from paraffinized blocks of tumor and “healthy” tissues. Relative copy numbers of 8 genetic loci (c-Myc, CDK12, CDKN1B, PRPF19, ERBB2, DDB1, GAB2, COL6A3) was assessed by RT-qPCR technique. Relative copy quantitation of a genetic locus was calculated as 2-ΔCt. The dose of the locus studied was considered equal to diploid set (2n) if RCQtumor/healthy was about 1. If RCQtumor/healthy was > 1.5 or < 0.5, then the locus dose was considered increased (≥ 3n) or decreased (≤ 1n), respectively.Results: For all genetic loci, an increase of relative copy quantitation in the ovarian tumor cells was observed compared to that in “healthy” tissues. There was a significant (рc-Myc (р = 0.001), DDB1 (р = 0.002), PRPF19 (р = 0.0001), and CDKN1B (р = 0.001). We identified differential thresholds for these genes that made it possible to predict an unfavorable disease course in the patients (р < 0.05). The strongest association with the risk of adverse outcomes was found for increased copy number of PRPF19 (odds ratio (OR) 7.3; р = 0.0001) and c-Myc (OR 6.8; р = 0.001).Conclusion: In this study, we determined the prognostic value of 4 oncogenic drivers, namely, DDB1, PRPF19, CDKN1B, and с-Myc, whose increased copy number was associated with an adverse disease prognosis in ovarian cancer patients.

Purpose of the study. Was to assess diagnostic informative value of liquid-based cytology optimized with genetic methods for the differential diagnosis of precancerous and malignant diseases of the cervix.Materials and methods. The study included 381 patients. Cervical pathologies were diagnosed with liquid-based cytology only and liquid-based cytology optimized with genetic methods of assessing the expression of miRNA‑20a, miRNA‑375, miRNA‑21 and –23b. Results of liquid-based cytology and genetic methods were verified by histological examination of the material. Statistical analysis was performed using descriptive statistics methods with the calculation of the mean and standard error of the mean. The mean values were compared with the help of the Mann-Whitney test.Results. Diagnostic results of liquid-based cytology were consistent with histological results in 107 (73.8 %) of 145 cervical cancer (CC) patients, in 52 (57.1 %) of 91 patients with high-grade squamous intraepithelial lesions (HSIL), and in 30 (65.2 %) of 46 patients with low-grade squamous intraepithelial lesions (LSIL). Optimization of liquid-based cytology by assessing the expression of miRNA‑21 and miRNA‑23b in the cervical epithelium improved the diagnostic sensitivity of the method from 73.8 % to 80 %, and its specificity from 94.1 % to 97.9 %. The diagnostic sensitivity and specificity of liquid-based cytology for differential diagnosis of CC and HSIL was 87 % and 78.8 %, respectively. Optimization of liquidbased cytology by assessing the expression of miRNA‑20a and miRNA‑375 in the cervical epithelium for the differential diagnosis of CC and HSIL improved the diagnostic sensitivity of the method from 87 % to 95.1 %, and its specificity from 78.8 % to 93.9 %.Conclusions. We revealed the most informative pairs of miRNAs in the cervical epithelium, as an analysis of their expression expanded the possibilities of liquid-based cytology both as a method for diagnosing CC and as a method for the differential diagnosis between CC and HSIL.

Purpose of the study. Studying possible management of postoperative pain and normalization of adaptation status in patients with reproductive system onychopathology using xenon- oxygen therapy.Patients and methods. The study included 97 patients receiving surgical treatment for reproductive cancer at National Medical Research Centre for Oncology in 2016–2020. All patients were divided into the main and control groups depending on the performed corrective therapy with xenon- oxygen mixture. Inclusion criteria were: established diagnosis — endometrial cancer, cervical cancer or ovarian cancer at the surgical stage of antitumor treatment, absence of decompensated concomitant pathology. In addition to general clinical tests, the intensity of symptoms in patients was assessed using a standardized Edmonton questionnaire, adaptation status and data of a numerical rating scale of pain were recorded, the severity of endogenous intoxication was measured using calculated lymphocytic, leukocyte, nuclear and leukocyte shift index. Stages of the study included functional assessment of the patient's condition before surgical treatment and on the 1st and 5th days of the postoperative period.Results. An analysis showed statistically significant differences between the groups: in the group of patients receiving xenon- oxygen therapy, 12.9% of patients complained of pain, while in the control group — 34.2%, on mild exertion 17.1% and 39.9%, respectively (pConclusion. The effectiveness of the chosen therapy with xenon- oxygen mixture demonstrates the possibility of anesthesia and normalization of the adaptive status of oncogynecological patients who underwent surgical treatment for reproductive cancers.

Aim. To analyze the expression of the p53 and Ki67 markers in patients with stage IIIC-IV ovarian cancer by immunohistochemical (IHC) method as a criterion for the treatment effectiveness. Materials and Methods. The study included 93 patients with ovarian cancer stage IIIC-IV aged 34-77 years. The patients were divided into 4 groups: group 1 – with neoadjuvant chemotherapy and intramuscular injections of recombinant interferon-gamma; group 2 – with neoadjuvant chemotherapy and intraperitoneal injections of recombinant interferon-gamma; group 3 (controls) – standard chemotherapy without interferon-gamma; group 4 (comparison group) – with surgery as the first stage of treatment. All tumor samples were studied by IHC method for the expression of p53 protein and Кi-67 marker of proliferative activity. Results. Neoadjuvant chemotherapy did not affect the levels of p53, compared to the group without chemotherapy. Inclusion of interferon-gamma to neoadjuvant chemotherapy resulted in statistically significant decrease in p53 expression, mainly with intraperitoneal administration. Study of proliferative potential of tumors in patients with intraperitoneal injections of interferon-gamma showed 1.3 times higher predomination of tumors with low proliferative activity com-pared to patients with intramuscular administration, 3.5 times higher compared to the group re-ceiving chemotherapy without interferon-gamma, and 7 times higher than in patients with sur-gery. High proliferative activity was noted only in tumor cells of patients without neoadjuvant chemotherapy. Conclusions. The IHC study demonstrated that inclusion of interferon-gamma to chemotherapy decreases proliferative potential of tumors and expression of р53 by tumor cells in ovarian cancer patients, especially with intraperitoneal introduction in the course of neoadjuvant chemotherapy.

Background: To predict the development and recurrence of cervical cancer (CC), we selected three oncoassociated miRNAs: miRNA-20a, -21, whose overexpression leads to the development of tumors, and -23b, which acts as an oncosuppressor. Aim: To evaluate the microRNA profile in the cervical epithelium for predicting CC recurrence in patients who underwent early treatment.Materials and methods: In the study of the informativeness of expression included 145 patients with T1a1-T2a1N0M0 CC who were followed up for 2 years after treatment. Expression of microR-NA-20a, -21 and -23b was analyzed in tumor tissue samples.Results: The risk of recurrence decreased from 1.0 to 0.92 after 1 year of the follow-up, and to 0.84 after 2 years. The initial expression of microRNA-20a and -21 in the cervical epithelium in patients with recurrent CC was 44% and 47% higher, respectively, than in patients without recurrence, while microRNA-23b expression was 46% lower. When initial levels of microRNA-20a and -21 expressions were 1.08 and 1.18, respectively, the risk of CC recurrence during the first two years after the surgery increased by 10.15 and 7.62 times, respectively. MicroRNA-20a expression in cervical epithelium equal to 1.08 was associated with 23% risk, and equal to 1.4 - with 79.7% risk. MicroRNA-21 expression equal to 1.18 was associated with 15% risk of CC recurrence; equal to 1.4 - with 55.5% risk; equal to 1.7 - 94.6%. Logistic regression showed that recurrence risks increased sharply when microRNA-23b expression declined.Conclusion: We registered higher levels of mi-croRNA-20a and -21 expressions and lower mi-croRNA-23b expression in patients with recurrent CC, compared to favorable course of the disease. An analysis of the expression profiles of micro-RNA-20a, -21 and -23b after CC diagnosis allow prognosis of recurrence risks within 2 years after the tumor removal surgery.

Omental malignant metastases are one of the challenging issues in oncology. The article provides a review of recent studies on the structure and functions of immune compartments of the greater omentum and their characteristic features that promote or inhibit tumor dissemination. The cellular composition of lymphoid nodules and milky spots is described, and functional and phenotypic properties of macrophages and lymphocytes are shown. Unique subpopulations of immunocompetent cells typical of this particular organ, as well as produced cytokines, are characterized. Particular attention is paid to the visceral adipose tissue surrounding immunocompetent cells and its effect on their functions. Analysis of the literature data has revealed a dual role, i.e. both protective and immunosuppressive one, of lymphoid structures of the greater omentum. The former is apparently associated mainly with a response to bacterial pathogens, while the latter is realized in cancer metastasis. The article focuses on immunological mechanisms that create local conditions for the growth and development of metastases, in particular, proinflammatory cytokines, chemokines, growth factors secreted by immune, tumor and mesothelial cells, and on the importance of the surrounding visceral adipose tissue for this process. The multidirectional prognostic significance of some local cellular and cytokine factors in cancer metastasis to the omentum and peritoneum is demonstrated. Possible approaches to treatment involving immunotherapy should be aimed at both elimination of tumor cells and overcoming the immunosuppressive environment. In this regard, reprogramming of macrophages, correction of hypoxic microenvironment, and the search for new control points seem promising.

e17532 Background: Vulvar cancer is a rare gynecologic tumor accounting for 2–5% of all gynecologic cancers. The epithelial-mesenchymal transition (EMT) is considered one of the factors contributing to the aggressive behavior of vulvar squamous cell carcinoma. E- and N-cadherins are key regulators of this transition. The purpose of our study was to identify the features of the expression of EMT markers in vulvar cancer as prognostic factors. Methods: The study included 30 patients aged 39-70 years, mean age 52 years, with vulvar cancer: control group (n = 15) with vulvar cancer T1-2N0-M0 (G2); main group (n = 15) with vulvar cancer T1-2N1-M0 (G2) with metastases to inguinal lymph nodes. Immunohistochemical study was performed on sections from paraffin blocks of tumors using monoclonal mouse antibodies to E-cadherin (12#, Cloud-Clone Corp.) and N-cadherin (389, Invitrogen) and the UltraVision Quanto Detection System HRP DAB. Expression of E- and N-cadherin was semiquantitatively analyzed according to the cell percentage and stain intensity: 0 (0-10%); 1+ (11-30%); 2+ (31-70%); 3+ (> 70%). Expression was considered positive if the score was ≥2. Pearson's χ² test and Spearman's correlation coefficient were used for statistical analysis. Results: The maximum staining for E-cadherin was observed in the control group (in 73%), for N-cadherin - in the main group (in 59%). Median expressions of E- and N-cadherin differed: in the control group, 70 [40-78] and 26 [0-40]; in the main group, 47 [45-60] and 59 [27-65]. Pearson's chi-squared test showed that patients with E-cadherin+ tumors (p = 0.006) statistically significantly prevailed in the control group, and patients with N-cadherin+ tumors (p = 0.027) prevailed in the main group. However, when considering the correlation between E-cadherin and N-cadherin in the groups, no statistically significant differences were found (rs = -0.042 in the control group and rs = 0.250 in the main group at p > 0.05), apparently due to the limited sample. Conclusions: The immunohistochemical study revealed a tendency towards loss of E-cadherin and increased expression of N-cadherin, which is often associated with poor prognosis.

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.

e15077 Background: Plant metabolites are traditionally a valuable source of anticancer drugs. The huge variety of molecular structures found in plants makes it possible to isolate substances that possess not only antimitotic, but also anti-migratory properties on cancer cells. The objective of this study was to evaluate the anti-migratory activity of selected metabolites from Petasítes sp. plants. Methods: Cells of cancer cell cultures PC3 (prostate cancer), A431 (skin cancer), CaCo-2 (colorectal cancer), HeLa (cervical cancer) and T98G (glioma) were seeded 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 the test substances at a concentration of 40 µM were added, and a wound healing assay was performed according to the standard procedure. The study included the following metabolites: 1) substance 1 (S1) - 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one; 2) substance 2 (S2) - 5-(hydroxymethyl)furan-2-carbaldehyde; 3) substance 3 (S3) - corynan. The wound area was measured using a Lionheart FX imager (BioTek, USA) at starting time point and after 48 hours of cultivation. Data are presented as mean±95% confidence interval (n = 12). Results: The studied cultures were found to be differently sensitive to the tested substances. Cells of CaCo-2 and PC3 cultures did not demonstrate a significant decrease in mobility, as the degree of scratch overgrowth did not decrease compared to the control without the addition of substances (α = 0.05, df = 22) in all variants of the experiment. However, the A431 skin cancer culture showed reduced motility in all three trials compared to the no-substance control. Namely, the wound area reduction was 81.52 ± 9.6% in the control, 63.77 ± 8.4% in S1 variant, 62.42 ± 6.3% in S2 variant, and 12.05 ± 7.1% in S3 variant. Also, a slowdown in cell motility was observed in HeLa (28.92±5.1% compared to 37.0±6.3% in control) and T98G cells (82.41±9.1% compared to with 97.35±1.9% in control) when incubated with S1. Taking into account the correction for multiple comparisons, only the results for A431 turned out to be significant, but the results obtained for HeLa and T98G cultures were not significant at a corrected significance level (α = 0.003, df = 22). Conclusions: Further studies of 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one anti-migratory properties are of particular interest, since it showed promising results in three of the five studied cancer cell cultures.

e21562 Background: Study of the influence of a pharmacological substance dihydrobromide-2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo-[1,2-a] benzimidazole on the tumor growth on a model of experimental transplantable B16 melanoma in mice. Methods: B16 melanoma strain were transplanted subcutaneously to 80 female C57/Bl6 mice weighing 18-20 g. Intragastric dihydrobromide-2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo-[1,2-a] benzimidazole injections (0.5 ml a day) began 48 h after tumor transplantation once a day for 10 days at single doses of 50, 220 and 500 mg/kg (groups 1, 2 and 3, respectively). Mexidol was used for comparison, and saline as control (injected at similar regimens and doses). Results: Intragastric enoxifol injections did not significantly inhibit the melanoma growth. Only group 2 showed inhibition of the tumor growth by 48.7 % on day 22. Median survival of mice in all experiment groups was similar to that in the control group. The number of metastatic nodes in the lung tissues in groups 1, 2 and 3 decreased by 2.1, 4.8 and 2.5 times, respectively, compared to controls. The average index of metastasis inhibition was 71.3 % in all groups being 1.3 times higher than in the mexidol group (54.2 %). Conclusions: The revealed significant changes in parameters of the metastatic activity of the tumor suppose that dihydrobromide-2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo-[1,2-a] benzimidazole at all studied doses has a greater effect on the development of spontaneous lung metastases and metastasis rates than on the growth of primary B16 melanoma in mice.

e15003 Background: Taxanes and platinum compounds are widely used in cervical cancer chemotherapy. In particular, a combination of 75 mg/m2cisplatin and 175 mg/m2 paclitaxel, which suggests a doses ratio of 1:2.3, is recommended as first-line systemic chemotherapy. Although this combination has performed well in clinical practice, little is known about its effect on cervical cancer cell biology in vitro. This study assessed the effect of combined application of cisplatin and paclitaxel in a dose ratio of 1:2.3 on HeLa cells. Methods: HeLa cells were seeded at 3000 cells per well in a 96-well plate (Eppendorf, Germany) and incubated for 24 hours in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA) at 37°C in an atmosphere containing 5.0% CO2. After 24 hours, the culture medium was replaced with a medium containing chemotherapy drugs in combination or as standalone agents in a series of two-fold dilutions: 0.03 - 32 µg/mL cisplatin and 0.07 - 73.6 µg/mL paclitaxel. After 2 h exposition to studied compounds the medium was replaced and cells were incubated for another 72 hours. At the end of the experiment, the number of living cells was assessed using the MTT test. Viability was determined as the number of living cells in % of the control without the addition of chemotherapy drugs. Results: The combination of cisplatin with paclitaxel demonstrated significantly greater efficacy than pure cisplatin in the concentration range from 0.03 μg/ml to 8 μg/ml. The difference between the mean viability values for each concentration between pure cisplatin and its 1:2.3 mixture with paclitaxel was significant at α = 0.05 (df = 14). The decrease in viability is especially dramatic at non-toxic concentrations of cisplatin (0.03 - 0.5 μg/ml) and is about 60% over this concentration range. However, when moving to toxic concentrations of cisplatin (0.5-32 μg/ml), the difference in the viability of HeLa cells between pure cisplatin and its combination with paclitaxel linearly decreases with increasing cisplatin concentration, becoming insignificant in the range of 8-32 µg/ml. However, when comparing the dose-response curves for pure paclitaxel and its combination with cisplatin, we see a significant reduction in cytotoxic activity compared with pure paclitaxel. The difference in cell viability reaches its maximum (45%) at a paclitaxel concentration of 2.3 µg/ml and is significant at α = 0.05 (df = 14). Conclusions: The combined use of cisplatin and paclitaxel in a dose ratio of 1:2.3 on HeLa culture has a greater cytotoxic activity compared to pure cisplatin. However, compared with pure paclitaxel, the effectiveness of its combination with cisplatin is significantly lower, which indicates the antagonism of the two substances.

e17500 Background: Our purpose was to analyze the rates of polymorphic allelic variants of genes of hemostasis system and methionine exchange in patients with female reproductive tumors. Methods: The study included 51 patients with histologically verified gynecologic tumors (group 1), including 28 patients (group 1a) with malignant tumors (cervical cancer (CC) n = 8, ovarian cancer (OC) n = 8, endometrial cancer (EC) n = 8, other cancers n = 4) and 23 patients (group 1b) with benign tumors, and 47 women without tumors (group 2). 12 polymorphic loci were studied by RT-PCR in genomic DNA samples: F2 (G20210А, rs1799963), F5 (G1691A, rs6025), F7 (G10976A, rs6046), F13 (G226A, rs5985), FGB G(-455)A (rs1800790), ITGA2-α2 (C807T, rs1126643), ITGB3-b (Т1565С, rs5918), PAI-1 4G(-675)5G, rs1799889), MTHFR (С677Т, rs 1801133 and A1298C, rs1801131), MTR (А2756G, rs1805087), MTRR (A66G, rs1801394). Groups 1, 1a and 1b were compared with controls (p1) and among themselves (p2). Results: The ratio of genotype frequencies maintained in the Hardy-Weinberg equilibrium in all gene loci except F7 (G10976A) in group 1 (p = 0.03). An alternative allele in the F2 gene was found only in group 2 (1.1%). The frequency of an alternative allele in the F5 gene in group 1 was 2.9%, including 1a – 1.8%, 1b – 4.3%, group 2 – 2.1%; F7 – 16.7%, 14.3%, 19.6% and 17.0%; F13 – 23.5%, 23.2%, 23.9% and 34%; FGB – 26.5%, 25.0, 28.3% and 25.5%; ITGA2 – 53.9% (p1= 0.03, OR = 1.89 (1.07-3.33), 48.2%, 60.9% (p1= 0.01, OR = 5.21 (1.22-5.17) and 38.3%; ITGB3 – 13.7%, 10.7%, 17.4% and 16.0%; PAI-1 – 47.1% (p1= 0.03, OR = 0.53 (0.30-0.93), 46.4%, 47.8% and 62.8%; MTHFR (Т) – 28.4%, 30.4%, 26.1%, 34.0%; MTHFR (С) – 34.3%, 28.6%, 41.3% (p1= 0.04, OR = 2.17 (1.02-4.61) and 24.5%; MTR – 18.6%, 19.6%, 17.4% and 27.7%; MTRR – 63.7%, 71.4%, 54.3% and 62.8%, respectively. TT genotype at the ITGA2-α2 (C807T) locus was more frequent in group 1 than in group 2 (23.5% vs 19.1%, p1= 0.01, OR = 6.54 (2.61-16.40); CT genotype was more frequent in group 1a than in group 2 (67.9% vs 38.3%, p1= 0.004, OR = 3.40 (1.27-9.13), and more frequent in EC than in group 2 (87.5% vs 38.3%, p1= 0.03, OR = 11.28 (1.28-99.40). GG genotype at the MTRR (A66G) locus was more frequent in group 1a than in group 1b (53.6% vs 26.4%, p2= 0.042). 5G5G genotype at the PAI-1 4G(-675)5G locus was more frequent in group 1 than in group 2 (31.4% vs 10.6%, p1= 0.04, OR = 3.84 (1.28-11.53), and more frequent in OC than in group 2 (75% vs 11%, p1= 0.0001, OR = 25.50 (3.96-160.20). AA genotype at the F7 (G10976A) locus was more frequent in CC patients than in group 2 (31.3% vs 17%, p1= 0.03, OR = 15.33 (1.20-195.75). Conclusions: Carriage of the AA genotype at the F7 (G10976A) locus may increase the risk of developing CC, and the CT genotype at the ITGA2-α2 (C807T) locus may increase the risk of EC. On the contrary, the alternative 4G allele at the PAI-1 4G(-675)5G locus was less common in patients with malignant tumors, especially OC, than in the group without cancer.

e17549 Background: The question of the mechanisms of platinum resistance development in advanced ovarian cancer (OC) remains open. The aim of the study was to reveal the relationship between the proliferative activity of OC cells, the disruption of apoptosis processes and the changes in the activity of transporter proteins that provide the efflux of chemotherapy drugs. Methods: Patients with advanced OC (T3-4N0-3M0-1) of postmenopausal age, sensitive (SPtPs, n = 23) and resistant to platinum drugs (RPtPs, n = 17), operated on after 3-6 cycles of neoadjuvant chemotherapy, were retrospectively studied. The parameters of proliferative activity (ki67), apoptosis disturbances (p53), as well as the expression of BCRP and Pgp transporter proteins were evaluated in OC tissue using immunohistochemistry methods. Statistical analysis was performed using the Mann-Whitney and Pearson tests (χ2), as well as the Spearman's rank correlation coefficient (SC). Results: In total, the dominance of the RPtPs over the SPtPs was noted in terms of intensity of the studied indicators (p < 0.05-0.003), the most significant for Pgp expression. At the same time, moderate and high values of these indicators were observed in SPtPs in 42-61% of cases, which reduced the predictive value of the revealed differences. It was of interest that statistically significant correlations between the indicators in SPtPs differed from those in RPtPs. Thus, the following values of the SC were observed in SPtPs: +0.848 (ki67-p53), -0.675 (ki67-BCRP), -0.575 (p53-BCR). In RPtPs, another intensity and/or direction of these relations were noted: +0.521 (ki 67-p53), +0.500 (ki 67-BCRP), +0.705 (p53-BCR). There were no statistically significant relationships between ki67-Pgp expression in SPtPs and RPtPs. The direction and intensity of the studied relationships could indicate changes in cell regulation in primary OC in SPtPs compared to RPtPs. Thus, in SPtPs, they reflected a significant contribution of apoptotic disturbances to the enhancement of OC cell proliferation, which was limited by the negative feedback mechanism via BCRP activity suppression indicated by the presence of an inverse correlation ki67-BCRP. In RPtPs, this mechanism was apparently already lost, which resulted in a change in the direction of the statistical relationship ki67-BCRP from reverse to direct one. Under these conditions, the positive contribution of apoptosis disruption to the proliferative activity of OC cells could obviously be supplemented by a synergistic increase in the activity of the BCRP transporter protein, which removes platinum drugs from OC cells. Conclusions: Patients with advanced OC may develop platinum resistance due to discoordination of proliferation, apoptosis, and regulation of the activity of the BCRP transporter protein.

e15002 Background: Bleomycin is a glycopeptide antibiotic with anticancer properties. Clinical data indicate a relatively low efficacy of bleomycin in mono regimen, which is also accompanied by side effects from high doses. That’s why so it is more often used in relatively low doses in combination with other chemotherapy drugs. In particular, in cervical cancer, bleomycin is used in combination with cisplatin in a dose ratio of 0.8:1. Despite good clinical results, it is not known how the two drugs interact in vitro. The aim of this study was to evaluate the effect of combined application of bleomycin and cisplatin on HeLa cells. Methods: HeLa cells were seeded at 3000 cells per well in a 96-well plate (Eppendorf, Germany) and incubated for 24 hours in DMEM medium (Gibco, USA) supplemented with 10% FBS (HyClone, USA) at 37°C in an atmosphere containing 5.0% CO2. After 24 hours, the culture medium was replaced with a medium containing chemotherapy drugs in combination or as standalone agents in a series of two-fold dilutions: 0.03 - 32 µg/mL cisplatin and 0.012 – 12.8 µg/mL bleomycin. After 2 h exposition to studied compounds the medium was replaced and cells were incubated for another 72 hours. At the end of the experiment, the number of living cells was assessed using the MTT test. Viability was determined as the number of living cells in % of the control without the addition of chemotherapy drugs. Results: The combination of cisplatin with bleomycin demonstrated significantly greater efficacy than pure cisplatin in the concentration range from 0.03 µg/ml to 8 µg/ml, and pure bleomycin in the range of 0.012 - 12.8 µg/ml. The difference between the mean viability values for each concentration of pure bleomycin or pure cisplatin and their 0.8:1 mixture was significant at α = 0.05 (df = 14). Observed decrease in the viability of HeLa cells in comparison with pure substances is most pronounced in the range of non-toxic concentrations for both substances in mono regimen, namely 0.03 - 0.5 μg/ml for cisplatin, and at concentrations less than 0.125 μg/ml for bleomycin. Here, the increase in the difference between the dose-response curves grows as the concentration of the studied drugs increases. At cisplatin concentrations of 0.5–32 μg/ml, the difference in HeLa cell viability between pure cisplatin and its combination with bleomycin, on the contrary, decreases linearly with increasing cisplatin concentration, becoming insignificant above 16 μg/ml. Conclusions: The combined use of bleomycin and cisplatin on HeLa culture in a dose ratio of 0.8:1 exhibits synergistic properties that are most pronounced in the concentration ranges of both substances that do not have a cytotoxic effect in mono regimen.

e17554 Background: Thrombosis in patients with gynecologic cancers worsens the outcome of antitumor treatment and is one of the leading causes of their death. The kallikrein-kinin system (KKS) is involved in both the regulation of thrombus formation and the development of cancer. The purpose of this study was to analyze characteristics of the KKS components in the blood in patients with endometrial cancer (EM) and ovarian cancer (OC) with and without secondary thrombosis (VTEC). Methods: The study included 39 patients, mean age 58.0±4.2 years: main groups – patients with OC T1c-3cN0M0 (n = 10) or EC T1a-2N0M0 (n = 9) with VTEC; comparison groups - patients with OC (n = 10) or EC (n = 10) without VTEC. EC was represented by adenocarcinomas (G1-G3), OC by serous carcinomas (90%) and clear cell adenocarcinomas (10%). The control group included healthy women of the corresponding age (n = 10). Blood levels of kallikrein 1 (K1), kallikrein 14 (K14), and kininogen (KG) were measured by ELISA after the surgery. Results: EC patients with VTEC showed 1.5 times (p < 0.05) higher blood levels of K1, compared to donors, while other indices were unchanged. EC patients without VTEC had 4 times lower KG levels, compared to donors and EC+VTEC. In OC, regardless of the presence or absence of VTEC, levels of K1 in the blood increased, as well as in women with EC+VTEC, by 1.5 times (p < 0.05) compared with donors, which was combined with a twofold increase in KG in OC+VTEC and a decrease in KG by 1.4 times in OC patients without VTEC. Blood levels of K14 increased only in OC patients without VTEC by an average of 1.9 times (p < 0.05) compared with donors and OC+VTEC. Conclusions: The revealed changes in some KKS components in the blood demonstrate the similarity (K1 increase) and differences (KG increase only in OC) in the pathogenesis of thrombosis associated with gynecologic cancers. The mechanisms of protection against VTEC in the early postoperative period also showed common (KG decrease) and specific features associated with the characteristics of cancer (K14increase only in OC). The development of therapy aimed at correcting the identified disorders will allow an antitumor treatment program for this category of patients with maximum efficiency improving their quality of life and survival.

e21568 Background: Despite the wide range of modern drugs for cancer drug therapy, there is still a problem of their high toxicity, which leads to the search for antitumor agents that selectively suppress or inhibit the growth of neoplastic cells. Over the past decades, many benzimidazole derivatives have been reported with potent antitumor activity due to its structural similarity to naturally occurring nucleotides. The aim is to study the effect of dihydrobromide-2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo-[1,2-a]benzimidazole on the growth and metastasis of transplantable experimental tumors after intraperitoneal administration. Methods: Melanoma B16 and Lewis lung carcinoma (LLC) were inoculated subcutaneously into 96 female C57/Bl6j mice (weighing 18–20 g). Intraperitoneal (0.3 ml/day) administration of the drug was carried out 48 hours after tumor inoculation 1 time per day for 10 days in single doses of 6.15 (1st), 30.75 (2nd) and 61.5 (3 -i) mg/kg. The control group of animals was injected with saline. The increase in life expectancy (T/C,%), the degree of tumor growth inhibition (TGI,%) and the metastasis inhibition index (MII,%) were calculated. Results: With intraperitoneal administration of 2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo[1,2-a]benzimidazole at a dose of 30.75 mg/kg, it increased the lifespan of animals with B16 (T/C = 151.3%). On the 7th day after the end of treatment in the 1st (6.15 mg/kg) and 2nd groups (30.75 mg/kg) a slight decrease in the volume of the primary tumor B16 was found, which remained by the 14th day only in 2 -th group (TGI = 23.6%). Also for LLC in the 30.75 mg/kg dose group, the T/C index was 171.6%. The volume of the primary subcutaneous tumor node was already 2.3 times lower on the 1st day after the end of treatment compared to the control (p < 0.05) and this effect persisted up to the 14th day (the tumor volume was reduced by 2. 1 and 1.9 times on the 7th and 14th days, respectively, at p < 0.05). In groups with B16, the use of the substance reduced the number of metastases in the 1st and 2nd groups by 2.5 and 3.0 times, respectively (p < 0.05), relative to the control. The frequency of metastasis was in the 1st group B16 -60.4%, and in the 2nd - 74.5%. Similar results of antimetastatic activity were shown for LLC: the number of metastases in the 1st and 2nd groups was reduced by 2.1 and 2.6 times, respectively (p < 0.05) compared with the control. MII was higher for LLC compared to B16 and amounted to 63.1 and 79.6 in groups for groups 1 and 2, respectively. Conclusions: When administered intraperitoneally, 2-(3,4-dihydroxyphenyl)-9-diethylaminoethylimidazo[1,2-a]benzimidazole dihydrobromide showed an antitumor effect against primary Lewis lung carcinoma and its lung metastases, as well as against lung metastases for melanoma B16 mice.

e17528 Background: Photodynamic therapy (PDT), with its high efficiency and selective impact on tumor cells without damaging healthy tissues, is an organ-sparing treatment preserving the anatomy and function of the cervix. Preservation of the reproductive function is the most significant criterion for the efficacy of an organ-sparing treatment. The purpose of this study was to evaluate the efficacy of PDT for early cervical cancer. Methods: PDT results were studied in 74 patients aged 18-40 years with early cervical cancer. The patients were divided into 3 groups depending on the type of transformation zone (TZ), the degree of invasion and the tumor site: group 1 - 36 patients with preinvasive cancer in the exocervix (TZ type I-II); group 2 - 34 patients with preinvasive cancer in the endocervix (TZ type III); group 3 - 4 patients with microinvasive cancer T1a1N0M0 with invasion of no more than 1 mm as an alternative to surgical intervention. Infection with high-risk genotypes of HPV (16, 18, 31, 33, 35, 45, 56) was detected with PCR in 62 (84%) women. All patients received PDT with the semiconductor Latus laser up to 3 W, a single-use diffusing fiber for the exocervix irradiation and a single-use cylindrical diffusing fiber for tumors in the cervical canal. Chlorin e6 was used as a photosensitizer. The efficiency criteria included the normalization of the colposcopic and morphological picture, and the HPV elimination confirmed by the PCR test. 4 to 8 procedures were required to restore the normal layer of stratified squamous epithelium. Results: PDT was followed by direct photocoagulation necrosis caused by the destruction of cellular structures, and ischemic necrosis as a result of damage and destruction of the microvasculature of the initial tumor changes in the cervical epithelium. Epithelialization of the cervix was completed by days 35-40. Normal morphological picture was observed in 96% of patients in group 1 and in 95% of patients in group 2; all patients in group 3 had a pronounced positive response. PCR 3 months after PDT showed a positive HPV reaction in 5.1% of patients. No negative changes in the cytogram were detected 6 and 12 months after PDT. The maximum follow-up period has lasted for 5.1 years. No relapses of the disease have been registered. During the follow-up period, four patients with preinvasive cancer and one patient with microinvasive cancer successfully gave birth to healthy children. Conclusions: PDT is an alternative treatment for precancerous and early malignant tumors of the cervix; it preserves the anatomical and functional integrity of the organ, which is important for the female reproductive function. The results of the study allow recommending PDT in the treatment of early cervical cancer.

e17548 Background: About 10-15% of malignant ovarian tumors (OC) are associated with hereditary diseases, and about 65-85% of hereditary OC have a mutation in BRCA1/2 genes. The aim of the study was to analyze BRCA1/2 genes hereditary mutation spectrum in patients in the South of Russia diagnosed with OC. Methods: We used EDTA venous blood and FFPE-blocks of 324 patients with histologically verified epithelial OC (T1-3cNx-1Mx-1, stage I-IV). BRCA1 gene mutations (185delAG, 300T > C, 2080delA, 4153delA, 5382insC, 3819delGTAAA, 3875delGTCT) and BRCA2 gene mutations (6174delT) were determined by real-time PCR. Results: The most common types of OC were poorly differentiated serous carcinoma (high degree of malignancy) - 75.6%, well-differentiated serous carcinoma (low degree of malignancy) - 13%, mucinous tumors - 2.8%, endometrioid tumors - 4.3%, clear cell adenocarcinomas - 4.3%. BRCA1/2 genes mutations were found in 52 (16%) patients with OC. The frequency of detected BRCA1/2 genes mutations was: 5382insC - 53.8%, 4153 delA - 9.6%, 300T > G - 21.2%, 2080delA - 7.7%, 185delAG - 3.8%, 6174delT - 3.8%. 3819delGTAAA and 3875delGTCT mutations were not found in the BRCA1 gene. All carriers were confirmed to have the same mutations in the tumor. There were no cases of sporadic changes in BRCA1/2. BRCA1 mutation carriers prevailed in the group of high-grade serous carcinoma patients (11.1%), where all cases of BRCA2 gene mutations were identified. In the group of low-grade serous carcinoma patients, the mutation rate was 1.5%, in the endometrioid adenocarcinomas group - 0.6%, in the mixed epithelial tumors group - 2.8%. In the group of patients with mucinous tumors, mutations were not detected. Conclusions: Thus, frequency of BRCA1/2 gene mutations in OC patients living in the South of Russia was 16.0%. Distribution of mutation types in the BRCA1 gene, with a predominance of 5382insC (53.8%) and 300T > G (21.2%), corresponded to their occurrence ratio in the European populations. BRCA1/2 gene mutations were registered more often in the group of high-grade serous carcinoma patients.

e17621 Background: An imbalance of sex hormones in endometrial cancer (EC) towards hyperestrogenism together with progesterone deficiency is considered to stimulate proliferation and suppress endometrial apoptosis. EC with squamous metaplasia is characterized by a more severe clinical course and the lack of an adequate response to standard treatment. The purpose of this study was to analyze the local hormonal profiles of tumors with squamous metaplasia in patients with obesity. Methods: The main group included 20 EC patients with squamous metaplasia, and control group included 35 patients with endometrioid adenocarcinoma: T1-3N0-1M0, mean age 64±3.2 years. Intact endometrial tissues were obtained during surgical treatment of patients with uterine fibroids (n = 20). All patients gave their written informed consent for the study. Levels of estradiol (E2), estrone (E1), testosterone (T), progesterone (P4), estrogen receptors ERα and ERβ, progesterone receptor RP4 and androgen receptors RA were measured by ELISA in samples of tumor and peritumoral (PT) tissues. Statistical processing of results was performed using the Statistica 10. Results: Tumor tissues of patients with EC, regardless of the histological structure, showed higher levels of estrogens and androgens and their receptors, compared with the intact endometrium, without significant changes in levels of progesterone and its receptor. In the main group, compared with the control group, E1 levels in tumors were lower by 1.6 times, E2 by 1.3 times (p < 0.05), but T higher by 2.4 times, ERα by 1.7 times, ERβ by 1.5 times (p < 0.05). PT, compared with the intact endometrium, had lower levels of E1 and E2 by 2 times and 1.5 times, P4 by 5.1 times, and T was lower only in PT with squamous metaplasia by 2.8 times. Levels of all steroid hormone receptors in PT were higher, regardless of the histological type of tumors. In PT of the main group, levels of E1 were 1.3 times lower (p < 0.05) than in controls, P4 – by 2.2 times, T – by 2.8 times, RP4 – by 1.7 times and ERβ by 1.8 times (p < 0.05), but ERα was 3.2 times higher. Conclusions: Tumors with squamous metaplasia, compared with endometrioid adenocarcinoma, are characterized by an imbalance of steroid hormones towards the predominance of androgens, and ERα plays a dominant role among estrogen receptors. The tumor probably synthesizes sex steroids independently and also pumps them over from the surrounding area. At the same time, malignant endometrial tumors, regardless of their histological structure, are more saturated with estrogens, androgens and their receptors, compared with the intact endometrium.

Цель. Оценить результаты применения неоадъювантной химиоиммунотерапии у пациенток с неоперабельными формами рака яичников. Методы. В исследование включены 72 пациентки с раком яичников стадии IIIС-IV, которые были разделены на три группы: с неоадъювантной химиотерапией и неоадъювантной химиоиммунотерапией с внутримышечным и внутрибрюшинным введением интерферона гамма. При оценке эффективности лечения изучали и сравнивали непосредственные и ближайшие результаты лечения. Дополнительно в свежем послеоперационном материале методом ДНК-проточной цитометрии проведены определение плоидности и анализ клеточного цикла. Выполнена оценка иммунного статуса до лечения и после окончания неоадъювантной терапии. Результаты. После проведения курсов неоадъювантной полихимиотерапии больным в группе с внутримышечным введением интерферона гамма выполнена операция полного объёма у 81,8% женщин, в группе с внутрибрюшинным введением интерферона гамма - у 87,5% пациенток (р ≤0,05), в контрольной группе (без применения интерферона гамма) операция полного объёма выполнена у 34,6% больных. Также выявлено, что в группах с применением интерферона гамма частота возникновения рецидива в группе с внутрибрюшинным введением интерферона гамма составила 25% (р

Ovarian carcinoma is the leading cause of death from gynecological cancer. Aim of the study is to reveal the role of VEGF-C comparing to VEGF-А in the progression of ovarian cancer. Material and methods. Tissue samples obtained from 76 patients with epithelial ovarian cancer (serous cystadenocarcinoma: T1N0M0; T2N0M0; T3NxM0; T4Nx-1M0) and 47 patients with cystadenoma were studied. Histological control was performed in all cases. Ovaries of 20 patients obtained during the surgery for uterine myoma were used as the intact tissue. All patients were 50.9 ± 2.9 years old. Levels of the growth factor as VEGF-A and its receptor VEGF-R1, as well VEGF-С and its receptor sVEGF-R3 - were measured by ELISA with the use of standard test systems. Results. VEGF-А levels in cystadenomas and intact tissue were similar, while in cystadenocarcinomas VEGF-А was significantly higher at all stages of the tumor development. sVEGF-R1 receptor in cystadenomas was lower in comparison with the intact tissue, in the contralateral ovary in T1N0M0 and in the tumorous ovary in T4Nx-1M0. VEGF-С level was higher significantly in all tumors, in cystadenocarcinomas it was higher if compared to cystadenomas. Its increase in the contralateral ovary in T1N0M0 differed from other tissue values being average. sVEGF-R3 receptor increased significantly at the later stages of ovarian cancer - T3NxM0 and T4Nx-1M0; its level was low only in the contralateral ovary in T1N0M0, and the values in other tissues were similar to the intact ones. Conclusion. The results show a high rate of lymphatic vessel formation in benign tumors at all stages of the development of the malignant tumor. The significant increase in VEGF-C level in the contralateral (non-tumorous) ovaries, compared to the intact tissue, allows considering VEGF-C, along with VEGF-А, as a pathogenetic factor of ovarian tumor development.

e17575 Background: Analysis of the typical structure of age, sex and territorial patterns of HPV spread is of interest, since it allows assessing the potential impact of modern immunoprophylaxis. The purpose of the study was to analyze the prevalence and variety of HPV types in dependence on gender and age. Methods: The study included 424 patients (334 women and 90 men) under 45 years of age (228 women and 46 men) and over 46 years old (106 and 44, respectively). Vaginal and cervical swabs were examined in women and urethral swabs in men. DNA extraction was performed by Real-time PCR. Results: HPV DNAs were found in 150(35.4%) patients, including 115(34.4%) women and 35(38.9%) men (p > 0.05). HPV was less frequently detected in older women, and more frequently in older men. Thus, HPV DNAs were found in 85(37.3%) women < 45 years and in 30(28.3%, p = 0.068) women >46 years; in men – 14(30.9%) and 21(47.7%, p = 0.071), respectively. Some HPV types were more frequent in younger patients, compared to older ones: in 26(26.3%) and in 5(9.8%), p = 0.068) of HPV-positive patients, respectively. The most common HPV types were 16(46.7%), 52(16.0%), 56(12.7%), 31(10.7%); HPV16 dominated both in females - 50(50.4%) and males - 12(34.6%). After HPV16, most frequent types in women were: 31-12(11.3%) and 52-13(11.3%), 18-12(10.4%) and 56-119(, 6%); in men - 52-11(31.4%), 56-8(22.9%), 45-4(11.4%) and 18-3(8.6%) types. The total share of two HPV types with the greatest oncogenic potential (types 16 and 18) was 85(56.7%) of all HPV-positive patients, and in women it was higher than in men: 70(60.9) vs. 15(42.9%, p = 0.046), respectively. Conclusions: Simultaneous infection with several HPV types was statistically significantly more often detected in younger patients compared with the older age group among men. HPV16 was the most common type both in men and women; however, the rates of other HPV types differed. Cumulative frequency of types 16 and 18 was statistically significantly higher in women, compared to men.

e17530 Background: Despite the progress made in diagnosis and treatment, ovarian cancer (OC) is still the third most common genital cancer. Poor results of therapy for various forms of OC stimulate the search for fundamentally new approaches to the treatment. One of the main reasons for resistance to chemotherapeutic agents includes multidrug resistance (MDR) of tumor cells. The most characterized of its various mechanisms is the elevated activity of the ABC family protein - ABCB1 (Pgp or MDR1). The purpose of the study was evaluation of the expression of this ABC transporter as a predictive factor in platinum-containing chemotherapy in OC patients. Methods: 100 patients aged 29-79 years with advanced stage IIIC - IV OC with/without ascites were recruited between 2016 and 2020. Depending on the treatment results, patients were divided into 2 groups: group 1 (n = 59) - patients with platinum sensitivity; group 2 (n = 41) - patients with platinum resistance during the treatment or within 6 months after its completion. IHC analysis was performed using rabbit polyclonal anti-MDR1 antibodies (Affinity Biosciences) diluted 1:600 and the Reveal Polyvalent HRP-DAB Detection System. The percentage and the intensity of staining were assessed: 0, 1+ weak, 2+ moderate, 3+ strong. MDR1 expression was considered positive when staining was detected in more than 10% (cut-off) tumor cells with intensities of 2+ and 3+. Statistical analysis of results was performed in the Statistica 13.0 program (StatSoftInc., USA). Results: Patients with MDR1+ prevailed in group 2 (98%), and patients with MDR1- in group 1 (32%). The average expression of MDR1 in tumor cells in group 2 (64.0±7.0) was statistically significantly higher by 1.7 times (p = 0.003) than in group 1 (37.2±6.8) by the Mann-Whitney U-test. When distributed according to Pearson's χ2 criterion, positive MDR1 as a risk factor in the platinum-resistant group increased the risks of refractory to platinum therapy by 19 times (95% CI 2.4-148.8). Conclusions: The study demonstrated predictive significance of the MDR1 biomarker in platinum treatment in patients with ovarian cancer.

e17561 Background: Contribution of local inflammation to the course of endometrial cancer (EC) is of interest since it may aggravate the disease. Methods: The study included 25 women aged 62+1.6 years with histologically verified EC (group 1a), 29 women aged 48+8.4 years with uterine fibroids (UF) (group 1b) and 13 non-cancer women aged 42+2.3 years (group 2, controls). Vaginal swabs were taken prior to antitumor treatment. Relative expression of genes encoding the synthesis of IL1B, IL10, IL18, TNFA, TLR4, GATA3, and CD68 in comparison with the reference gene B2M was determined by Real-time PCR using the ImmunoQuantex kit (Russia). An integral parameter of an inflammation index was calculated using binary logistics, and the ratios of the listed indicators were also calculated. 12 parameters were analyzed in total. Groups Ia and Ib were compared with healthy people (p1) and with each other (p2). Results: 4 of 12 studied parameters in EC patients differed from control; no differences were found in UF patients. 7 statistically significant differences were registered between EC and UF. Relative expression of the gene encoding the synthesis of IL10 was maximal in EC (2.8+0.2 vs. 1.8+0.3 in healthy women (p1= 0.006) and 1.8+0.2 in UF (p2= 0.002)). The IL10/IL18 ratio in EC statistically significantly exceeded the ratios in UF patients and in healthy women (61.4+21.7 (p1= 0.003), 46.3+32.2 (p2< 0.001) and 53.7+47.1 respectively). The TNFA/IL18 ratio in EC patients was also higher than in UF patients (1.1+0.7 vs. 0.5+0.2 (p2= 0.035)) due to higher TNFA levels (3.7+0.1 vs. 3.3+0.1 (p2= 0.006)), one of which effects includes stimulation of neoangiogenesis. The ratio of TLR4/GATA3, on the contrary, was lower in EC than in UF (0.7+0.3 and 1.5+0.9 respectively (p2= 0.001). Apparently, it characterized antimicrobial immunity because both of these genes are involved in the response to PAMP (TLR4) and in the genesis of intraepithelial lymphocytes related to innate immunity, as well as in the development of a humoral response via stimulation of Th2 (GATA3). The latter, in addition, is involved in the ontogenesis some organs, including the vagina and uterus. The highest TLR4/GATA3 ratio was found in UF, which, in our opinion, indicated the predominance of the inflammatory process in UF and local immunosuppression in EC. The inflammation index determined with the ImmunoQuantex test system did not differ between the studied groups of women. Conclusions: Some differences were observed in local reactions of immunity and inflammation in benign and malignant uterine tumors, involving the prevalence of immunosuppressive and angiogenic factors in EC and inflammatory factors in UF.

e17527 Background: Resistance to chemotherapy is a complex problem in the treatment of patients with ovarian cancer (OC), and this phenomenon is associated, among other things, with the presence of cancer stem cells (CSCs). The purpose of the study was to determine the amount of CSCs in tumors of OC patients with different responses to chemotherapy they receive. Methods: Samples of ovarian tumors were obtained from 100 patients (aged 29-79 years) with advanced ovarian cancer stage IIIC-IV with/without ascites. All patients recruited in 2016-2020 received standard combination treatment with surgery and platinum-containing polychemotherapy (Pt CT). Based on the results of neoadjuvant Pt CT, patients were divided into groups: patients with stabilization or progression–group 1 “without effect”; patients with complete/partial regression–group 2 with positive effect. All patients gave their informed consent for the study. Cell suspension obtained from the tumor fragments was processed using monoclonal antibodies labeled with various fluorochromes: CD45-APC-Cy7, CD44-FITС, CD133–РЕ according to the manufacturer's instructions (BD, USA). The percentage of CSCs was determined using the FACS Canto II flow cytometer (BD, USA). The amount of cells with CSC markers (CD44+, CD133+, CD44+CD133+) was calculated as a percentage from the total amount of CD45--cells. Statistics: STATISTICA 13 (StatSoftInc., USA). Results were presented as Me (LQ; UQ). The significance of differences was assessed using the Mann-Whitney test (the differences were considered significant at p- _cells, as well as CD45-CD44+ cells, in OC tumors did not differ between patients of groups 1 and 2: 79.0 (72.1; 85.9) vs. 82.8 (76.7; 89.0), and 6.2 (3.3; 9.1) vs. 6.6 (3.5; 9.8), respectively. The percentage of cancer cells with the CD45-CD133+ phenotype in group 2 was significantly lower than in group 1: by 52% (p≤0.05), 3.2 (2.4; 3.9) vs. 6.7 (3.8; 9.6), respectively. The amount of CD45-CD44+CD133- CSCs in tumors in group 1 was lower than in group 2: by 82% (p≤0.05), 0.6 (0.3; 0.7) vs. 3.3 (1.8; 4.7), respectively. Conclusions: The results demonstrated certain differences in the distribution of CSCs in tumors of OC patients with different sensitivity to chemotherapy, which may be one of the factors determining the treatment outcomes.

e17503 Background: Early stages of cervical cancer (CC) can be treated with surgery or radiation therapy, metastatic CC is incurable, and therefore new diagnostic approaches based on highly effective molecular markers are required. As such markers, the Copy Number Variation (CNV) is of great interest. Purpose of the study: to analyze the peculiarities of genes CNV in normal cells, primary tumor and lymph nodes tumor metastases cells in patients with CC. Methods: Tissue sections from FFPE-blocks of 300 patients with CC were used for the study. Primary tumor, tumor metastatic and normal cells were isolated using non-contact laser microdissection (Palm MicroBeam, Carl Zeiss). DNA was extracted from the cells using the phenol-chloroform method. Determination of 14 genes CNV (LAMP3, PRKAA1, TORC2, FOXO3, HDAC5, MEF2C, MLXIPL, EP300, HNF4A, TP53, SREBF1, SREBF2, PPARGC1A, and CCND1) was performed by Real-Time qPCR method. Statistical analysis was performed using the Mann-Whitney test; Bonferroni's correction was applied to correct multiple comparisons. Results: An increase in the CNV of LAMP3 and TORC2 genes by 2.5 and 3.2 times (p < 0.05), respectively, and a decrease in the CNV of TP53 and FOXO3 genes by 2.3 and 2.0 times (p < 0.05), respectively, were found in the primary tumor cells relative to normal cells. The effect of the simultaneous change in the CNV of LAMP3, TORC2, TP53 and FOXO3 genes was observed in 60% of the sample, and LAMP3 and TP53 genes - in 80%. In cells of lymph nodes tumor metastases an increase in the CNV of CCND1 and TORC2 genes by 2.0 and 3.5 times (p < 0.05), respectively, was found, as well as a decrease in the CNV of PPARGC1A gene by 2.1 times (p < 0.05) relative to normal cells. A simultaneous change in the CNV of CCND1, TORC2, and PPARGC1A was observed in 70% of the sample, and CNV of CCND1 and TORC2 genes - in 85%. Primary tumor and metastases cells differed in the CNV of CCND1 (2.5 times higher in metastatic cells), LAMP3 (3.0 times lower in metastatic cells) and FOXO3 genes (2.0 times higher in metastatic cells). Conclusions: Thus, the primary tumor and lymph nodes metastases cells differ in the level of genes CNV from normal cervix cells. Accordingly, the CNV of LAMP3, TORC2, TP53 and FOXO3 genes may have the potential for diagnosing a primary cervical tumor, and the CNV of CCND1, TORC2 and PPARGC1A genes for diagnosing its metastases to regional lymph nodes.

e17508 Background: Cervical cancer (CC) is one of the main causes of death among patients with gynecological tumors. As a rule, patients die from relapses and metastases which rates reach 30-45%. According to research data, treatment of patients with distant metastases is ineffective, and half of them die within 9.7 months, while the average survival is 15.9±0.26 months. The development of many pathological processes is considered to be associated with an increase in free-radical reactions leading to oxidative damage to various biomolecules. The purpose of the study was to analyze some characteristics of the free-radical oxidation and antioxidant defence in metastasizing CC. Methods: The study included 56 patients aged 29-73 years with stage IIB-IV CC after antitumor treatment. The main group – 27 patients who developed metastases within 4 months to 8 years after the CC diagnosis; controls – 29 patients with non-metastatic CC; donors – 19 healthy women aged 27-61 years. The accumulation of carbonyl derivatives in blood plasma proteins was detected in the reaction with 2, 4-dinitrophenylhydrazine. The induced oxidative modification of proteins was stimulated with Fenton's reagent. Free-radical processes was evaluated by the intensity of peroxide-induced luminol-dependent plasma chemiluminescence and the content of nitric oxide metabolites; the intensity of lipid peroxidation - by the content of malondialdehyde (MDA); the activity of catalase and ceruloplasmin was also studied. Results: CC progression was accompanied by increasing lipid peroxidation and spontaneous oxidation of blood plasma proteins. MDA levels in patients with metastases increased by 57.8% compared to donors and by 34.3% compared to patients without metastases (p < 0.05). The concentration of 2, 4-dinitrophenylhydrozones increased on average by 4 times compared to donors and by almost 2 times compared to patients without metastases. Patients with metastasizing CC demonstrated elevated levels of products of the interaction of nitric oxide and its derivatives with proteins and peptides - 3-nitrotyrosine and nitrosoglutathione, compared to both donors (by 31.4% and 55.3%) and patients in remission (by 38.1% and 34.5%). Chemiluminescence activity increased by 54.5% compared to donors (p < 0.05) and by 93% (p < 0.01) compared to controls. Catalase activity in the blood plasma of patients with metastases increased by 54.1% compared to donors, but was lower than the values in the control group (by 22.1%). Ceruloplasmin activity was increased only in patients without metastases (by 33%). Conclusions: The process of CC metastasis is accompanied by a greater intensity of oxidative processes of both proteins and lipids, as well as depletion of the adaptive capabilities of the body's antioxidant system.

e15010 Background: Ovarian cancer (OC) is the fourth most common cancer in women aged 40-54 years. Despite advances in cancer research, no existing methods provide effective early diagnosis and treatment of OC, so the search for new medications is an urgent task. The effectiveness of the XAV-939 pharmacological substance, which inhibits the Wnt signaling pathway, was studied in an orthotopic patient-derived xenograft of human ovarian cancer. Methods: A PDX model was created in 20 female immunodeficient Balb/c Nude mice. A tumor fragment was implanted into the ovary, and the ovary was transposed under the skin to better visualize the xenograft growth. When the average tumor volume was 67.4 mm3, animals were divided into 4 groups: group 1 – controls (n = 5); group 2 – animals receiving paclitaxel (n = 5); group 3 – animals with XAV-939 (n = 5); group 4 – animals with combined paclitaxel+XAV-939 (n = 5). The control group received the carrier substance, and the test substances were administered at 10 mg/kg; the substances were administered intraperitoneally twice a week for 22 days. The tumor growth was assessed throughout the experiment by measuring its linear dimensions (length and width), and the volume and tumor growth index were calculated. The data were statistically processed using Microsoft Excel and STATISTICA 10. The Mann-Whitney nonparametric test was used to assess the differences between the control and experimental groups. Results: No statistically significant differences were observed in the indices of tumor growth between groups 2 (paclitaxel) and 1 (control), and groups 3 (XAV-939) and 1 (control). However, the difference was statistically significant between groups 4 (paclitaxel+XAV-939) and 1 (control). The tumor growth inhibition index was calculated at the end of the experiment. It was maximal in group 4 (paclitaxel+XAV-939) – 67.78%; in groups 2 (paclitaxel) and 3 (XAV-939), it was 21.39% and 30.33%, respectively. Conclusions: An orthotopic PDX model of ovarian cancer was generated, and the activity of XAV-939 towards the OC xenograft was studied; the greatest efficacy of the preparation was noted in combination with paclitaxel.

e17560 Background: The purpose of the study was to analyze the structure and activity of the blood flow in the preserved ovaries after gynecologic surgeries. Methods: We monitored 50 patients (mean age 35.8±0.5 years) using sonography after ovary-sparing hysterectomy for benign, precancerous and preinvasive malignant uterine diseases. For revascularization, the preserved ovaries were attached to the lateral wall of the small pelvis or to the base of round ligaments of the uterus. Results: Ovarian hypertrophy was recorded in 3 women during the first year of the follow-up; the ovaries returned to the initial size without correction by 24 months. By the third year of observation, 94% of women had normal gonadal sizes, and only 2 women had ovarian hypoplasia. During the entire observation period (3 years), in 91% of cases, we could visualize the regular maturation of follicles in the preserved ovaries, which indicated the preservation of ovulatory function. The use of Doppler techniques during 24 months of the follow-up demonstrated that the blood circulation in the attached ovaries after hysterectomy was decreased on the average by 20.3%, compared to the intact ovaries. Conclusions: After the critical period of hysterectomy, the blood flow in the ovaries in almost all patients remains sufficient during the first 2-3 years, stimulating the regular growth of follicles and maintaining the gonadal activity. The quality of life of the operated patients during the first three years of follow-up remains optimal without additional correction with hormone replacement therapy.

e17528 Background: Resistance to chemotherapy is a complex problem in the treatment of patients with ovarian cancer (OC). Various intercellular intermediates are involved in the phenomenon, and cytokines are the most important of them; however, the involvement of individual representatives in the processes is poorly understood. Our purpose was to analyze levels of some cytokines in tumors of OC patients with different responses to chemotherapy they receive. Methods: Samples of ovarian tumors were obtained from 100 patients (aged 29-79 years) with advanced ovarian cancer stage IIIC-IV with/without ascites. All patients recruited in 2016-2020 received standard combination treatment with surgery and platinum-containing polychemotherapy (Pt CT). Based on the results of neoadjuvant Pt CT, patients were divided into groups: patients with stabilization or progression–group 1 “without effect”; patients with complete/partial regression–group 2 with positive effect. All patients gave their informed consent for the study. Levels of IL-6, IL-8 and IL-10 were measured in cytosolic fractions of tissue samples using standard ELISA test systems (Vektor-Best, Russia). Results were presented as Me (LQ; UQ). The significance of differences between the samples was assessed using the Mann-Whitney test (the differences were considered significant at p

e17516 Background: The combination treatment with plasmapheresis and non-specific immunotherapy for locally advanced cervical cancer (CC) is shown to be effective. We studied the effect of immunotherapy with dendritic cell vaccine (DCV) as a part of combination therapy on the intensity of the lipid peroxidation (LPO) and activity of antioxidant enzymes. Methods: Plasma and erythrocyte content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD), catalase, glutathione dependent enzymes and the content of reduced glutathione (GSH) in erythrocytes were determined in 24 patients with CC of T2a-2bN0-1M0 stages and in 25 women without cancer. The patients underwent neoadjuvant chemotherapy (CT) with cisplatin and bleomycin, followed by 2 cycles of DCV, for a total of 6 courses of CT and 12 cycles of DCV. Results: Initially, patients demonstrated intensification of free radical processes enhancing after PCT and the first two DCV: MDA in erythrocytes elevated by 52.2-111.7%, in the blood plasma by 2.2-3.5 times. Subsequent DCV cycles normalized the LPO. MDA levels after 4 DCV cycles were lower than the initial values in erythrocytes by 30.4% (p = 0.04), in plasma by 55.6% (p = 0.007), i.e. were similar to the levels in healthy women. The SOD activity was decreased by 16.7-27.3% (p < 0.005) after 4-6 DCV cycles and normalized after the 7th cycle. The catalase activity in erythrocytes was decreased before treatment and after PCT by 42.4% (p = 0.000000), during vaccine therapy by 20.5-37.7% (p < 0.0005). In reduced catalase activity, inactivation of hydrogen peroxide was realized, apparently, by glutathione peroxidase (GPO), and its increased activity (by 37-102.3%, p≤0.00005) was supported by a high GSH content - by 11.6-52.4% (p < 0.02) above the norm at all stages after the 2nd DCV cycle. And only with the complete normalization of the erythrocyte MDA content, the GPO activity after the 7th DCV cycle decreased by 46.7% (p = 0.000000) relative to the levels before the treatment and did not differ significantly from the values in donors. Conclusions: Vaccine therapy helped to enhance the activity of the glutathione system becoming the most important component of antioxidant protection in the blood of CC patients. Already 3-4 DCV cycles normalized the LPO sharply increased in CC patients during CT. Multi-course vaccine therapy allowed maintaining the optimal oxidative status of the blood.

5592 Background: Photodynamic therapy (PDT) is an effective method for the treatment of various cancers resulting in apoptosis, autophagy and ischemic necrosis of irradiated tissues. The purpose of the study was to analyze the efficacy of PDT for pre-invasive vaginal cancer treatment. Methods: PDT results were studied in 20 patients aged 32-65 years with verified pre-invasive vaginal cancer. All patients received PDT with the Latus diode laser and Photolon or Photolan photosensitizers. The effect was evaluated with extended colposcopy. The criteria for efficiency included normalization of the colposcopic picture and the absence of atypical cells. The sizes of the irradiation fields varied from 1.5 to 2 cm, the number of fields - from 1 to 4, the power density - from 0.1 to 0.17 W/cm2, the light dose - from 40 to 100 J/cm2. The duration of a PDT session varied from 10 to 30 min, depending on the number of irradiation fields. The irradiation field necessarily included an area of normal tissue 3-5 mm surrounding the lesion. 4 to 6 sessions were required to restore the normal layer of stratified squamous epithelium. The antitumor efficacy of PDT was evaluated based on the results of visual observation of changes in the area of the treated pathological foci and information on the presence or absence of clinical symptoms of the disease 1 and 3 months after the treatment (WHO criteria). Results: Complete regression was registered in 100% of patients after 3 months. Repeated courses of PDT were required in cases with a wide spread of pathological foci and the impossibility of their simultaneous irradiation. At follow-up after 1 month, 3 of 20 patients (15%) showed local foci of atypical changes in the epithelium managed with repeated PDT courses. In 6 months, stable remission of the disease clinical symptoms in the treated pathological foci was registered. The results of the cytological study performed 3 months after PDT were normal in 100% patients; no negative changes were registered 6 and 12 months after PDT. Conclusions: The results of PDT in the treatment of patients with pre-invasive vaginal cancer demonstrated its high therapeutic efficacy and a minimal number of adverse reactions, which allows recommending PDT in the treatment of pre-invasive vaginal cancer.

e17535 Background: Analysis of the nature of cancer cell release and interaction with the microenvironment determines the fundamental basis of ovarian cancer pathogenesis, which is necessary for the development of new methods for peritoneal dissemination treatment and prevention. The purpose of the study was to analyze levels of VEGF-A, VEGFR1, VEGF-C, VEGFR3, TGF-β, IGF-1, IGF-2 and IGFBP3 in omental tissues (O) with and without metastasis (Mts) in ovarian cancer patients. Methods: Samples of O tissues were obtained from 51 ovarian cancer patients aged 60±1.9 years (G2-G3 serous cystadenocarcinoma – AC, Т2-3NxM0-1). 17 non-cancer patients of similar age were controls (C). Levels of VEGF-A, VEGFR1, VEGF-C, VEGFR3, IGF-1, IGF-2, IGFBP3 and TGF-β1 were measured in Mts and O tissues by standard ELISA test systems. Results: VEGF-A levels in O of AC patients were 2.6 times higher than in C. In Mts, VEGF-A and VEGFR1 exceeded the levels in C (by 6.3 and 3.1 times) and O (by 2.5 and 2.8 times, respectively). VEGF-C in O was lower than in C by 5.5 times, while VEGFR3 was 2.2 times higher. In Mts, VEGF-C and VEGFR3 exceeded the levels in C (by 7.8 and 3.6 times) and O (by 43.3 and 1.6 times, respectively, p

e17574 Background: The purpose of the study was to assess vaginal biocenosis in patients with gynecological cancers (GCs) and in non-cancer women. Methods: The study included 50 patients aged 52.3+2 years with histologically verified GCs (cervical cancer – 23, endometrial cancer – 27) – group 1, and 22 non-cancer women aged 42.4+2.3 – group 2. Vaginal swabs were taken prior to antitumor treatment. Real-time PCR was used to determine the total bacteria mass (TBM) and DNAs of 16 groups of microorganisms: Lactobacillus spp., Enterobacteriaceae, Streptococcus spp., Staphylococcus spp., Gardnerella vaginalis+Prevotella bivia+Porphyromonas spp., Eubacterium spp., Sneathia spp.+Leptotrichia spp.+Fusobacterium spp., Megasphaera spp.+Veillonella spp.+Dialister spp., Lachnobacterium spp.+Clostridium spp., Mobiluncus spp.+Corynebacterium spp., Peptostreptococcus spp., Atopobium vaginae, Candida spp., Mycoplasma hominis, Ureaplasma (urealyticum+ parvum), Mycoplasma genitalium. Results: Vaginal biocenosis was assessed as normocenosis in 12 (24%) women in group 1 and 12 (54.5%, p = 0.012) women in group 2; conditional normocenosis - 2 (4%) and 3 (13.6%, p > 0.05); dysbiosis - 36 (72%) and 7 (31.8%, p = 0.0017), respectively. Anaerobic bacteria prevailed in women with dysbiosis in both groups (46% and 22.7%, p = 0.005); aerobic dysbiosis was registered in 16% and 4.5% (p > 0.05), mixed dysbiosis – in 10% and 4.5% (p > 0.05) of women of groups 1 and 2, respectively. Normal flora ( Lactobacillus spp.) was observed in 23 (46.0%) in group 1 and 17 (77.3%, p = 0.02) in group 2. Streptococcus spp. >10% of TBM were found in 9 (21.9%) women in group 1 and not found in group 2 (p = 0.049). Gardnerella vaginalis+Prevotella bivia+Porphyromonas spp. >10% of TBM were found in 27 (54.0%) in group 1 and 6 (27.3%, p = 0.031) in group 2; Staphylococcus spp. - 13 (26.0%) in group 1 and 12 (54.5%, p = 0.031) in group 2. Conclusions: Development of gynecological tumors was accompanied by a sharp decrease in Lactobacillus spp., a significant increase in Gardnerella vaginalis+ Prevotella bivia+ Porphyromonas spp. and Streptococcus spp., vaginal dysbiosis with a predominance of anaerobic and mixed microflora.

e17573 Background: Our aim was to evaluate ALDH1 expression as a stem cell marker in primary and recurrence vulvar squamous cell carcinoma. Methods: 27 patients aged 27-80 years with vulvar squamous cell carcinomas, together with their verified histopathological and clinical data were included in the study. All of them underwent treatment the first stage of which was surgery in National Research Centre for Oncology in 2015-2020. In 10 of them I stage of vulvar squamous cell carcinomas was diagnosed, in 13 – II stage, 4 had relapses after complex treatment and 2-10 years` remission. All paraffin-embedded samples of the primary and recurrent vulvar carcinoma were recruited. The presence of ALDH1 was detected by immunohistochemistry using murine monoclonal antibodies to ALDH1 (clone B-5 Santa Cruz Biotechnology) diluted 1:800 and Reveal Polyvalent HRP-DB Detection System. Positively stained cells were counted among all the tumor cells as well as a part of tumors positive for ALDH1 ( > 10% of cells). Statistics analysis was performed by program STATISTICA 13.0 (StatSoftInc., the USА). Results: By immunohistochemical staining, the expression of ALDH1 was observed in 5 patients with I stage of vulvar squamous cell carcinomas, in 5 patients with II stage and in all the patients with recurrence. Mean values of ALDH1 expression in patients with I stage were 14.1±4.3%, in patients with II stage – 16.0±3.6%, in patients with recurrence – 29.6±3.6%. Maximal ALDH1 accumulation was observed in tumor cells of patients with recurrence, it was 1.9 times higher than in the II stage (p = 0.03) and 2.1 times exceeded ones of the I stage (p = 0.027). All the differences were statistically significant according to Student`s t-test (p < 0.05). The was no statistically significant difference between values of the patients with I and II stages (p = 0.738), but it is noteworthy that ALDH1-expressing cells prevailed in tumor stroma especially in II stage. Conclusions: There is some discrepancy in the assessment of ALDH1 prognostic value in vulvar carcinoma patients as well as in understanding if it is related predominantly to cancer stem cells or to normal stem cells either. Our results though obtained on a few cases of vulvar squamous cell carcinomas show that ALDH1 expression in recurrent carcinomas markedly exceeds that in primary tumors and is related to tumor cells while in primary tumors is related rather to stromal cells.

e18022 Background: Locally advanced and progressive cervical cancer is always accompanied by the pain syndrome. The common analgesics include non-steroidal anti-inflammatory drugs and opioids. The purpose of the study was to assess allogeneic dendritic cell vaccine (DCV) as an alternative analgesic in patients with advanced cervical cancer. Methods: The pain syndrome dynamics was assessed in 20 patients with advanced T3-4N1M0-1 cervical cancer receiving subcutaneous paravertebral injections of allogeneic DCV in a total alternating dose of 5-10 million cells every 2 weeks. Patients received DCV for one year, with a total of 24 vaccine therapy sessions and a total dosage of 180 million dendritic cells. The pain intensity was assessed on a verbal rating scale: 0 – no pain; 1 – mild pain; 2 – moderate pain; 3 – severe pain; 4 – extremely intense pain. Results: Prior to the therapy, 15 patients (75%) had severe pain; 2 (10%) - moderate pain; 3 (15%) - extremely intense pain. After 4-6 DCV injections, the pain intensity decreased, patients refused opioid analgesics. After 10-12 DCV sessions, 19 (95%) (p < 0.05) women had no pain at all, patients denied additional pain relief with non-opioid analgesics. Unrelieved pain was registered only in one cervical cancer patient. Conclusions: DCV injections in patients with advanced cervical cancer provide pain relief thereby improving their quality of life.

e18029 Background: Numerous studies on cervical cancer confirm an important role of specific genomic changes in the onset and development of cervical intraepithelial neoplasia and their effect on the progression of cervical cancer. Solid tumors are characterized by genomic changes leading to a change in the DNA sequence copy number. The purpose of the study was to reveal changes in the relative copy number of the ESR1, ESR2, GPER1, STS, SULT1A1, SULT1E1, CYP1A1, CYP1A2 genes responsible for the reception and metabolism of estrogens in cervical tissues in endophytic and exophytic patterns of tumor growth in order to find predictive markers of malignancy. Methods: The study included 40 patients aged 28-65 years with cervical cancer of endophytic (n = 20) and exophytic (n = 20) growth patterns. Eligibility criteria included a morphologically confirmed cervical squamous cell cancer T1b-2aN0M0, stage I-II. The Thermo Scientific GeneJET FFPE DNA Purification Kit was used for the DNA extraction from FFPE blocks of tumor and healthy tissues. DNA concentrations were measured on the Qubit 2.0 fluorimeter (Invitrogen, USA) using the Quant-iT dsDNA High-Sensitivity (HS) Assay Kit (Invitrogen, USA). Results: The relative copy number of the GPER1, SULT1A1, CYP1A1 genes in tumor samples increased (p < 0.05) compared with normal tissues in the total sample of patients diagnosed with cervical cancer. In contrast to the total sample, an increase in the SULT1A1 gene dosage did not reach a statistically significant level in any group (p = 0.242 and p = 0.157); the copy number of the GPER1 locus significantly increased only in the group with the endophytic growth pattern (p = 0.040), as well as the CYP1A2 gene dosage (p = 0.025). Patients of 36-55 years with endophytic tumors showed a statistically significant (p < 0.05) increase in the GPER1 and CYP1A1 gene copy numbers with the rates of 41.7% and 66.7%, respectively, as well as an increased amplification of the CYP1A2 gene in 41.67% of patients. In women of 56-75 years with endophytic tumors, an increase in the copy numbers of the ESR2, GPER1, SULT1A1 genes was observed with a frequency of 50%, 100% and 75%, respectively. Patients aged 20-35 and 36-55 years with exophytic tumors showed a statistically significant (p < 0.05) increase in the CYP1A1 gene copy numbers in 33.33% and 45.45%, respectively. Conclusions: The results suggest the use of the GPER1, SULT1A1 and CYP1A1 gene copy numbers as biomarkers of cervical tumors.

e18037 Background: Renal dysfunction in cancer patients is an urgent problem of clinical practice in oncology which can often challenge the efficacy of anticancer treatment and the disease prognosis. The purpose of the study was to reveal risk factors for renal dysfunction in oncogynecological patients. Methods: A prospective randomized study of the renal function included 174 patients aged 54-82 years with reproductive tumors. The control group included 34 patients with benign uterine tumors. Glomerular filtration rate (GFR) was estimated using the CKD-EPI equation. Results: GFR in the main group was 71.6±2.3 mL/min/1.73m2. Renal dysfunction was found in 42.5%. The majority of patients (115 patients, 66.1%) had concomitant diseases classified as modifiable risk factors for the development and progression of chronic kidney disease (KDIGO, 2017), including hypertonic disease in 46.6% (χ2= 4.4, p < 0.05), diabetes in 39.1% (χ2= 7.9, p < 0.05), cardiovascular diseases in 27.6% (χ2= 3.9, p < 0.05), and morbid obesity in 68.9%. This confirmed the decisive importance of hypercholesterolemia, hyperglycemia and hypertension as risk factors for reduced GFR. 52.9% of patients had a combination of these factors, which together with the initial renal dysfunction suggested a high probability of acute renal failure in the perioperative period. Conclusions: The assessment of kidney function in patients with gynecological cancers is required to determine the intensity of treatment in the perioperative period, the general disease prognosis and risks of possible complications.

6035 Background: Photodynamic therapy (PDT) is an effective treatment for various cancers ensuring maximum preservation of the viability of healthy tissues surrounding the tumor. The purpose of the study was to reveal the effectiveness of PDT in treatment for preinvasive cervical cancer. Methods: The study included 45 patients aged 22-53 years with preinvasive cervical cancer. The patients were divided into two groups depending on the type of the transformation area and the tumor site: group 1–on the exocervix (type I-II), n=24; group 2–on the endocervix (type III), n=21. Infection with high-risk genotypes of HPV (16, 18, 31, 33, 35, 45, 56) was detected with PCR in 37 (82%) women. All patients received PDT with the semiconductor Latus laser up to 3 W, a single-use diffusing fiber for the exocervix irradiation and a single-use cylindrical diffusing fiber for tumors in the cervical canal. Photoditazine and photolon were used as photosensitizers. Effectiveness criteria included the normalization of the colposcopic picture, the absence of atypical cells, and the pathogen elimination confirmed by PCR. Results: A normal cytogram profile was observed after PDT in 84% of group 1 and in 88% of group 2. PCR 3 months after PDT showed a positive HPV reaction in 9.1%. Neither group of patients had negative changes in cytogram after 6 and 12 months. Repeated HPV DNA tests detected HPV DNAs in 2.8% in group 1 and 3.2% in group 2. The effectiveness of PDT did not depend on the photosensitizer. The maximum follow-up period has lasted for 4.5 years, with no recurrences registered. During this period, three young women successfully gave birth to healthy children. Conclusions: PDT is an alternative treatment for pre-tumor and initial tumor pathology of the cervix with preservation of the anatomical and functional integrity of the organ, which is important for the female reproductive function. The results support the use of PDT in treatment for preinvasive cervical cancer.

e13088 Background: The development of breast cancer (BC) and ovarian cancer (OC), as well as their chemoresistance may be due to the presence of mutations in many genes, but most often in BRCA1/2 and CHEK2. Early diagnosis and subsequent preventive interventions improve the survival of patients with mutations in the BRCA1/2 genes. Genotyping of the gene inactivation events predicts a high sensitivity of BC and OC to platinum-containing cytostatics. Our purpose was to study the spectrum of germline mutations in the BRCA1/2 and CHEK2 genes in BC and/or OC patients in the South of Russia. Methods: The study included 622 patients of 10 nationalities with BC and/or OC from the South of Russia. Genomic DNA was isolated from peripheral blood leukocytes; mutations were detected by Real-Time PCR in the BRCA1 (185delAG, 300T > C, 2080delA, 4154delA, 5382insC, 3819delGTAAA, 3875delGTCT), BRCA2 (6174delT), CHEK2 (1100delC, IVS2+1G > A, 470T > C) genes. Results: Mutations in the BRCA1/2 gene were found in 13.5% of cases, in CHEK2 – 6.2%. In the BRCA1/2 gene, the frequency of 5382insC mutation was 76.2%; 4153 delA – 9.5%; 300T > G – 7.1%; 2080delA – 3.6%; 185delAG, 3875delGTCT, 6174delT - 1.2% each. In the CHEK2 gene: 470Т > C – 73.1%; IVS2+1G > A – 23.1%; 1100delC – 3.8%. Two patients showed a combination of 5382insC mutation in the BRCA1 gene and 470T > C mutation in the CHEK2 gene. The observed prevalence of mutations in BRCA1 was generally consistent with that for European countries (p = 0.062). Identification of 470T > C mutation in CHEK2 in two cases confirmed the final diagnosis of Li-Fraumeni syndrome (OMIM: 609265); however, we failed to establish a family history of cancer in one patient. A high-risk cancer group was formed for prophylactic purposes based on the data of genotyping for major mutations in the BRCA1/2 and CHEK2 genes in patients with BC/OC and their healthy relatives. Conclusions: The frequency of BRCA1/2 and CHEK2 mutations in the studied multinational population of Caucasian patients with BC and/or OC was 17.2%. The 5382insC BRCA1/2 (76.2%) and 470Т > C CHEK2 (73.1%) mutations were the most frequent which corresponded to the occurrence of these SNPs in populations of European countries.

e18079 Background: Immunotherapy in oncology has now proven effective, but standard approaches have not yet been defined. Ways to optimize and increase the effectiveness of treatment with immunomodulators, in particular interferon-gamma preparations, in such patients remain an urgent problem of gynecological oncology. The purpose of the study was to improve direct results of ovarian cancer treatment. Methods: The study included patients aged 50-77 years with inoperable ascitic ovarian cancer (verified by cytological examination of ascitic fluid), stage IIIC-IV, receiving neoadjuvant chemotherapy (CT) in combination with immunotherapy with interferon-gamma (IFNγ). Group I – 26 patients, CT with carboplatin (AUC-6), paclitaxel (175 mg/m2); group II – 22 patients, chemoimmunotherapy and intramuscular IFNγ (ingaron) - 500 000 IU on day 1, 1 million IU on days 2,3,5, a similar CT on day 4; group III – 24 patients, chemoimmunotherapy and intraperitoneal IFNγ (ingaron) - 500 000 IU on day 1, 1 million IU on days 2,3,5, a similar CT on day 4. Patients received on average 2-3 therapy cycles. Results: Progression was registered only in group I in 3.8%; complete response in patients without IFNγ – 7.8%, with intramuscular and intraperitoneal IFNγ – 27.3% and 37.5% (p≤0.05) respectively. Surgical treatment followed in all patients, with total surgeries in 87.5% of patients with intraperitoneal IFNγ. DNA cytometry showed the minimal number of aneuploid tumors in patients with intraperitoneal IFNγ (16.6%), while in patients without immunotherapy 38.4%. The DNA index statistically significantly decreased by 1.3 times in patients with intraperitoneal IFNγ compared with patients without IFNγ (1.11±0.01% vs. 1.4±0.05% respectively) (p≤0.05). Levels of CD3+CD4+ cells elevated by 1.2 times (from 36.2±1.6 to 44.9±3.78%, p≤0.05) in patients with intramuscular IFNγ; intraperitoneal IFNγ caused an increase in CD3+ lymphocytes by 1.3 times (from 62.1±2.8 to 77.9±2.94%, p < 0.05) and CD3+CD4+ by 1.4 times (from 36.2±1.6 to 50.6±5.9%, p≤0.05). Conclusions: Interferon-gamma (ingaron) in combination with CT improves direct results of the treatment; intraperitoneal injections of interferon-gamma demonstrated better outcomes and tolerance confirmed by immunological and DNA cytometric parameters.