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Introduction. Enzyme-linked immunosorbent assay of biochemical markers is one of the most important methods for diagnosing tumors. One of these markers is an inducer of expression of matrix metalloproteases EMMPRIN/CD147. Changes in its expression are associated with the progression of some tumors. This study is the first work devoted to the study of the content of the soluble form of the transmembrane glycoprotein EMMPRIN (sEMMPRIN) in the blood serum of patients with various bone tumors.Aim. To study the content of sEMMPRIN in the blood serum of patients with malignant bone tumors, its relationship with the clinical and morphological characteristics of neoplasms and prognosis.Materials and methods. The study included 88 patients with malignant tumors (osteosarcoma – 37 cases, chondrosarcoma – 39, chordoma – 5, Ewing’s sarcoma – 7) and borderline (11 cases) bone neoplasms, of which 14 patients were under the age of 18 years. The control group consisted of 29 healthy donors, 8 of which were under the age of 18 years. The concentration of EMMPRIN was determined in the serum of patients and donors with reagents for direct enzyme immunoassay “Human EMMPRIN” (R&D, USA) in accordance with the manufacturer’s instructions and expressed in nanograms (ng) per 1 ml of blood serum. The obtained data were processed using the GraphPad Prizm 9.4 program. When comparing indicators and analyzing their relationships, we used the nonparametric Mann–Whitney and Kruskal–Wallis tests. Overall survival was analyzed using the Kaplan–Meier method.Results. Our analysis of the sEMMPRIN content in the blood serum of patients with bone tumors did not reveal statistically significant differences between the control group and patients with borderline and malignant tumors, both in adults and in children. At the same time, a trend towards a decrease in the level of sEMMPRIN in the blood serum was noted in the presence of a malignant neoplasm of the bone compared with the corresponding control group. Additionally, we found that the content of sEMMPRIN is associated with age and higher in the group of patients under 18 years of age, both among healthy donors and oncological patients. An analysis of the association of sEMMPRIN content with clinical and morphological characteristics did not reveal statistically significant patterns, however, a trend towards an increase in the level of the marker with disease progression in both studied age groups was observed, which is consistent with other studies conducted on other solid tumors.Conclusion. ELISA revealed the marker sEMMPRIN in the blood serum of all examined children and adults with borderline malignant bone tumors and healthy donors. At the same time, the levels of sEMMPRIN did not differ between the above groups, however, there was a tendency for a decrease in the concentration of the marker in patients with bone sarcomas compared with the control group, regardless of age.

Background. Implementation of immunotherapy in clinical oncological practice has significantly improved the results of cancer treatment. It resulted in the need for seeking new markers to assess the effectiveness of therapy and the disease prognosis.Aim. To analyze the content of soluble forms of PD-1 and PD-L1 immune checkpoint proteins in the blood serum of patients with non-small cell lung cancer and esophageal squamous cell carcinoma and their association with clinical and morphological characteristics of the disease and the disease prognosis.Materials and methods. The study included tumor samples obtained from 43 patients with non-small cell lung cancer and 21 patients with esophageal squamous cell carcinoma. The concentration of sPD-L1 and sPD-1 in the blood serum was determined using enzyme-linked immunosorbent assay (ELISA). The Mann – Whitney test was used to determine statistically significant differences in independent groups. A correlation analysis was performed using the Spearman’s rank correlation coefficient. Overall survival was analyzed by constructing survival curves using the Kaplan – Meier method and a Cox proportional hazards model. The differences were considered statistically significant at p < 0.05.Results. The study showed that sPD-1 and sPD-L1 were found in the blood serum of both cancer patients and healthy donors, and their concentrations did not differ significantly. It was shown that the high concentration of sPD-L1 in the blood serum of patients with non-small cell lung cancer was significantly associated with the late stage of the disease and was an independent unfavorable prognostic factor. It should be noted that for patients with esophageal cancer, an unfavorable prognostic marker was the high concentration of the soluble form of PD-1 protein, and not PD-L1 ligand, as in case of lung cancer.Conclusion. The content of sPD-1 and sPD-L1 in the blood serum can have different prognostic significance for various types of cancer, and further studies are required to confirm their clinical usability.

Introduction. Carcinoid syndrome is the most common functional syndrome in patients with neuroendocrine tumors. More than 40 biochemical factors are responsible for the manifestation of carcinoid syndrome, among which serotonin is the most important. The study of biochemical markers of carcinoid syndrome and associated carcinoid heart disease is an important aim of laboratory examination in neuroendocrine tumors patients.Aim. Analysis of levels and diagnostic efficiency evaluation of chromogranin A (CgA), serotonin, pro-brain natriuretic peptide (proBNP) and platelet-derived growth factor (PDGF-BB) in the blood serum of neuroendocrine tumors patients with various clinical manifestations, including carcinoid syndrome and carcinoid heart disease.Materials and methods. 66 patients with neuroendocrine tumors of various localizations were examined (pancreas – 24 cases, small intestine – 21, large intestine – 6, lungs – 10, unkown primary focus – 5). 38 patients had liver metastases. In 43 patients, a clinic of carcinoid syndrome was observed, 16 had signs of carcinoid heart disease. The control group consisted of 30 practically healthy people. Serum levels of CgA, serotonin, and PDGF-BB were determined by enzyme immunoassay in microplate format: Chromogranin A NEOLISA (Eurodiagnostica, Sweden), Serotonin ELISA (IBL, German), and PDGF-BB ELISA Kit (Invitrogen, USA). The proBNP analysis was performed on a Cobas e601 automated analyzer (Roche, Switzerland).Results. In carcinoid syndrome, the medians of CgA, serotonin, and proBNP were the highest, differing statistically significantly from the control group. In patients with G3 tumors, the median PDGF-BB was statistically significantly higher than in controls, in contrast to G1 and G2. The highest diagnostic sensitivity in the general neuroendocrine tumors group was in CgA – 63.6 %, with a specificity of 100 %. In patients with carcinoid syndrome, the highest diagnostic sensitivity was characteristic of serotonin and chromogranin A (79 %), while in patients with CAD clinic, proBNP had the highest sensitivity – 93.8 %.Conclusion. The study revealed the high efficiency of СgA, with the highest sensitivity in common forms and tumors with high biological activity. Serotonin can be used in the diagnosis of carcinoid syndrome, associated with cardiofibrosis development. Pro-brain natriuretic peptide is a highly sensitive and specific marker of carcinoid heart disease. The highest levels of PDGF-BB are associated with a high grade of neuroendocrine tumors malignancy.

Introduction. Immunoenzyme assay of biochemical markers is one of the most important methods for examination of pa- tients with neuroendocrine tumors (NETs). Along with the generally accepted NET marker chromogranin A (CgA), another member of the granin family, chromogranin B (CgB), can serve as a complementary marker.Objectives. Analysis of CgB as an additional to CgA biochemical marker in the blood serum of patients with gastric and pancreatic neuroendocrine tumors.Materials and methods. We examined 79 patients with gastic (n = 14) and pancretic (n = 65) NETS, and 42 particularly healthy people, who were included in the control group. CgB and CgA were determined with ELISA method using the Human Chromogranin B (USCN, China) and Chromogranin A NEOLISA (Eurodiagnostica, Sweden) test systems., Results. CgB levels in gastric and pancreatic NETs were significantly higher than in control group. CgB concentrations were independent of tumor spread and its biological activity. ROC analysis in common group of NETs relative to control group showed AUC for CgB = 0.869 and for CgA AUC = 0.82. According to results in common group of NET patients when used isolated, CgA and CgB have comparable diagnostic sensitivity, which increases in complex use to 82.5 %. In the group of NET patients with low levels of CgA (15.7 ng/ml) was observed in 53.6 % of cases.Conclusion. The combination of CgB and CgA in gastric and pancreatic NETs could increase the diagnostic efficacy of bio- chemical diagnostics. The received data confirms the significance of CgB as a complementary biomarker of NETs.

B7-H3, also called CD276, is a type I transmembrane glycoprotein that is encoded on human chromosome 15. It was discovered back in 2001. The original study described it as a positive co-stimulant, as it can stimulate T-cell response and IFN-y production. However, recent researches have shown that B7-H3 is involved in T-cell inhibition. A B7-H3 receptor has not been yet identified, and this may explain the complex immunomodulatory activity of B7-H3, as it can have more than one binding partner with different functions. Expression of the B7-H3 protein has been found on activated immune cells such as T-cells, NK cells and antigen presenting cells. Interestingly, it is overexpressed in a wide range of tumor cells and is associated with disease progression and outcome. The soluble form of this protein is also of particular interest. Increased sB7-H3 levels in the plasma of bone tumor patients might be their important diagnostic criterion.

Background: Evaluation and search for new molecular markers of renal cell cancer, first of all, associated with angiogenic and invasive activity, continue to be highly relevant. In our previous publications, we have assessed the potential diagnostic value of matrix metalloproteinases (MMP) 2, 7, 8, and 9, their tissue inhibitor type 1 (TIMP1) and components of the VEGF signaling system in renal cell cancer. Aim: To assess the role of serum VEGF, VEGFR1, VEGFR2, MMP2, 7, 8, 9, and TIMP1 levels in renal cell patients as diagnostic and prognostic markers of overall survival. Materials and methods: 99 renal cell cancer patients (94 primary and 5 at progression) were recruited into the study. The control group included 97 healthy control blood donors. Ninety three (93) primary patients with renal cell cancer were followed for 1 to 45 (median, 26) months for assessment of their overall survival. Serum concentrations of the study proteins were measured by direct immunoenzyme analysis (Quantikine® ELISA kits, R&D Systems, США). Results: Serum VEGF, VEGFR1, VEGFR2, MMP7, MMP8, and TIMP1 levels in renal cell cancer patients are significantly higher than those in the control group. The diagnostic characteristics of the markers are considerably different, the most reliable marker with 84% sensitivity at 87.5% specificity being MMP7. VEGFR1, MMP7, MMP8, and TIMP1 were positively associated with disease stage and TNM indices. MMP7 and TIMP1 levels also increased with a higher tumor grade. MMP7 was found to be a significant unfavorable prognostic factor for overall survival: the 3-years survival in those with low (< 6.3 ng/ml) marker level amounted to 93%, whereas with high, 51% (p < 0.001). MMP7 prognostic value remained significant also in stage I renal cancer: after 3-years' follow-up, all patients with low MMP7 were alive, while survival of those with high marker levels was 72% (p=0.02). Increased serum MMP8 level (> 51 ng/ml) also had an unfavorable prognostic value in the whole renal cell cancer patient group, with 3-years' survival being 78 and 58% for low and high levels, respectively (p < 0.01). The components of VEGF signaling system, MMP2, MMP9, and TIMP1 had no significant prognostic values. Conclusion: MMP7 should be viewed as the most promising diagnostic and prognostic renal cell cancer marker. VEGF and its soluble receptors could be useful for monitoring of patients receiving anti-angiogenic treatments and prediction of their sensitivity to these agents.

Background. Tumors of stroma of the sex cord include a family of tumors that are diverse in structure and biological characteristics, including hormone-active ovarian tumors, such as granulosa cell tumor (GCT) of the ovary and a tumor from Sertoli–Leydig cells. For these types of tumors, of particular importance is the analysis of biochemical markers, among which the most promising is inhibin B.The objective of the study. Comparative analysis of inhibin B levels in the blood serum of patients with stromal cell tumors and other types malignancies.Materials and methods. 64 patients with primary ovarian tumors were examined: 31 – GCT of the ovary, 16 – tumors from Sertoli–Leydig cells, 17 – adenocarcinomas. Comparison group – 20 patients with malignant tumors of other localizations, control – 74 healthy women and 37 patients with benign ovarian tumors. Inhibin B was determined in blood serum using the standardized Inhibin B Gen II ELISA (Beckman Coulter, USA) immunoassay.Results. The analysis of inhibin B levels in last days of the luteal phase, show an increase of marker level in patients with GCT and tumors from Sertoli–Leydig cells, while in ovarian adenocarcinomas and malignant tumors of other locations inhibin B secretion doesn’t differ from the control. The sensitivity of inhibin B in diagnostics of GCT was 93.5 %, in tumor from Sertoli–Leydig cells – 81.3 % with specificity – 100 %.Conclusion. Inhibin B is an effective biomarker of GCT and ovarian tumors from Sertoli–Leydig cells, which results must be interpreted according the functional state of the ovaries.

Background: The utility of biochemical markers in the monitoring of treatment efficacy in patients with neuroendocrine tumors (NETs) goes beyond any doubt. However, there are still no clear criteria for the assessment of clinically significant abnormalities of the main NET biomarkers chromogranin A (CgA) and serotonin. Aim: To evaluate the value of serial measurement of serum CgA and serotonin in the monitoring of the treatment effect in NET patients. Materials and methods: Serum CgA and serotonin levels were measured in 107 patients with NETs at baseline and at 3–4 weeks after the end of treatment (chemotherapy, biotherapy), as well as in 60 healthy controls. We used enzyme immunoassay based on Chromogranin A NEOLISA (Euro Diagnostica) and Serotonin ELISA (IBL International GmbH) test systems. Results: There was an association between CgA levels and the efficacy of chemotherapy in NET patients. With progression of the disease, median CgA increased significantly from 412 to 2679 ng/mL (p = 0.012), whereas in the patients with partial response it decreased from 811 to 254 ng/mL (p = 0.023). The ROC analysis showed the 33% cut-off for significant CgA changes for progression, with sensitivity of 80.0% and specificity of 95.6%. A decrease (of more than 33% compared to baseline levels) or absence of significant CgA changes was associated with stabilization of the disease or with partial response to treatment. Significantly decreased CgA levels were found in 75.0% cases of partial response and 43.48% of stabilized patients, whereas the absence of any significant changes in 25 and 66.7%, respectively. There was no association between serotonin levels and the disease behavior under treatment.Conclusion: CgA could be used as a sensitive marker of NET progression on chemotherapy.

Rationale: We have previously identified a group of microRNA genes (MIR-107, MIR-1258, MIR-130b, MIR-34b/c, MIR-9-1, MIR-9-3 et al.), whose methylation was involved into the development and progression of ovarian cancer. Aim: To expand the range of microRNA genes hypermethylated in ovarian cancer and to study the role of this modification in the pathogenesis and progression of ovarian cancer. Materials and methods: The study was performed on a series of 76 ovarian cancer and 13 peritoneal metastases samples. The method of bisulfite DNA conversion followed by methylation-specific polymerase chain reaction (PCR) was used to assess the methylation status of the microRNA genes; the expression of these genes was measured by quantitative real-time PCR. Results: Compared to histologically unchanged ovarian tissue, there was a significant increase in methylation frequencies in the tumor samples for 6 microRNA genes studied: MIR-124-1, MIR-124-2, MIR-124-3, MIR-125B-1, MIR-127, and MIR-129-2 (p ≤ 10-3). The expression level of 4 microRNAs (miR-124-3p, miR-125b-5p, miR-127-5p, miR-129-5p) encoded by these genes was suppressed, with a significant correlation between changes in their expression levels and the gene methylation (rs = 0.63–0.94, p ≤ 10-4). In addition, there were statistically significant associations between methylation of 5 genes (MIR-124-2, MIR-124-3, MIR-125B-1, MIR-127, and MIR-129-2) and the parameters of cancer progression, such as its clinical stage, metastatic spread, tumor size and invasion, and to a lesser extent with a decrease in the differentiation grade. The association of 5 microRNA genes with metastatic spread was confirmed by the analysis of peritoneal macro-metastases from 13 patients. Conclusion: We have demonstrated the functional significance of aberrant methylation in a group of microRNA genes for suppression of their expression in ovarian carcinomas. There is an association of microRNA gene hypermethylation with the progression of ovarian cancer, including metastatic spread to the peritoneum.

Background: Ovarian cancer is one of the most common oncologic diseases holding the frst place in mortality related to neoplasms of female genitalia. Along with active surgical intervention, contemporary ovarian cancer treatment includes various chemotherapeutic regimens which in many cases are quite effective, but relapse and death rates still remain high. In the recent years, major attention has been paid to the possibility of ovarian cancer immunotherapy associated with the discovery of the so-called “immune checkpoint” signaling, i.e. programmed cell death-1 / programmed death-ligand 1 (PD-1/PD-L) pathway, controlling intensity and duration of autoimmune response at physiologic conditions. Tumor PD-1 and/or PD-L1 expression is being actively studied as a predictor of anti-PD-1/PD-L treatment efficacy; however, this approach has certain limitations and problems that might be probably bypassed by determination of soluble PD-1 (sPD-1) and its ligand (sPD-L1) in serum or plasma.Aim: Comparative evaluation of sPD-1 and sPD-L1 content in plasma of healthy women and of patients with benign or borderline ovarian tumors and ovarian cancer, as well as the analysis of associations between these markers and main clinical and pathologic characteristics of ovarian cancer.Materials and methods: Sixty two (62) patients with ovarian neoplasms aged 32 to 77 (median, 56.5) years were enrolled into the study. Fifteen (15) patients had benign tumors, 9 had borderline, and 38, ovarian cancer. The control group included 17 healthy women aged 24 to 67 (median, 49) years. Plasma sPD-L1 and sPD-1 concentrations were measured with standard enzyme immunoassay kits (Afmetrix, eBioscience, USA).Results: Plasma sPD-L1 and sPD-1 levels in ovarian cancer patients (median, 41.3 and 48.0 pg/ml, respectively) did not differ significantly from those in the control group (49.5 and 43.8 pg/ml). sPD-L1 level in the patients with benign tumors (median, 22.2 pg/ml) was signifcantly lower than in the control (p < 0.01). The lowest sPD-1 level in plasma was found in the patients with borderline ovarian neoplasms, the difference with the ovarian cancer group being statistically signifcant (p < 0.05). No correlations between sPD-L1 and sPD-1 plasma levels were found in any of the study groups. sPD-L1 level signifcantly increased with disease stage (R = 0.44; p < 0.01), the most signifcant increase being observed at the most advanced IIIC stage (p < 0.05 as compared to all other stages). sPD-L1 was also signifcantly higher in the patients with ascites than in those without ascites. Plasma sPD-1 concentration was not associated with the indices of ovarian cancer progression, though its median was 1.3–1.44 times lower in the stage I than in the stage II–III patients, and decreased in those with the tumor size above 10 cm (assessed by ultrasound examination) and in the patients with ascites. No statistically signifcant associations of the markers' levels with tumor histological type and differentiation grade of ovarian cancer were found.Conclusion: sPD-L1 level in ovarian cancer patients correlates with disease progression and can be considered as a promising marker for monitoring of anti-PD-1/PD-L1 treatment efficacy. Potential clinical implications of sPD-1 require further studies.

Objective: Mutations in TP53 lead to loss of function (LOF) or gain of function (GOF) of the corresponding protein p53 and produce a different effect on the tumor. Our goal was to determine the spectrum of somatic TP53 variants in BRCA1/2 associated high-grade serous ovarian cancer (HGSOC).Methods: The population under study comprised of HGSOCs with pathogenic variants in BRCA1 (n = 78) or BRCA2 (n = 21). Only chemo-naive and platinum-sensitive patients were included in this study. The case group of the IARC database (n = 1249) with HGSOC not stratified by BRCA status was used as a reference. A custom NGS panel was used for sequencing TP53 and mutational hot-spots of other genes, and p53 expression was evaluated by immunohistochemistry for 68 cases of HGSOCs.Results: Somatic TP53 variants (95) or inhibition of wild-type p53 expression (3) were observed in 98 cases. The sample with normal p53 had CDKNA1 variants. The frequency of truncating variants was significantly higher than in the reference cohort (30.3 vs. 21.0%, p = 0.01). Most of the samples (41/68) demonstrated low (or absent) expression of p53, and 17 samples overexpressed p53. LOH was typical for TP53 nonsense variants (14/15). In total, 68/95 samples were LOH positive and showed LOH in all tumorous cells, thus indicating the driver effect of TP53 mutations. Three specimens had KRAS, BAX, APC, and CTNNB1 subclones variants.Conclusion: High frequency of TP53 truncating variants, the low expression of mutant p53, and low incidence of oncogene mutations show potential GOF properties of p53 to be poorly represented in BRCA1/2 associated HGSOC.

Background: Over the last 10 years the incidence of gastric cancer has declined significantly. Nevertheless, it remains one of the most prevalent malignancies both in Russia and worldwide. Therefore, the problems of early diagnostics, prognosis and individualized treatment choice are still on the agenda. Much attention is paid to the evaluation of molecular biological characteristics of the tumor, as well as to the development of multiparametric prognostic systems for gastric cancer based on its identified characteristics. An important place among potential tumor biological markers belongs to matrix metalloproteinases (MMPs) involved into all the stages of tumor progression, first of all, into the regulation of invasion and metastasizing.Aim: Comparative quantitative evaluation of some MMP family members (MMP-2, 7, and 9) and one of the tissue MMP inhibitors (TIMP-2) levels in the tumors and adjacent histologically unchanged mucosa in gastric cancer patients, the analysis of their associations with the main clinical and pathological features of the disease and its prognosis.Materials and methods: Sixty six (66) primary gastric cancer patients (32 male and 34 female) aged 24 to 82 years (median, 61 year) were recruited into the study. Twenty two (22) patients were with stage I of the disease, 11 with stage II, 28 with stage III, and 5 with stage IV. The concentrations of the proteins studied were measured in the tumor and unchanged mucosa extracts by standard direct ELISA kits (Quantikine®, R&D Systems, USA).Results: Tumor MMP-2, 7 and 9 levels were significantly increased, compared to those in the adjacent histologically unchanged mucosa, in 80, 70 and 72% of gastric cancer patients, respectively, while the increase of TIMP-2 level found in 61% of the tumors was not statistically significant. Tumor MMP-2 and TIMP-2 content was increasing significantly with higher T index – size and advancement of the primary tumor (p < 0.01 and p < 0.05 respectively). Tumor MMP-2 level was also increasing in parallel with the N index (regional lymph node involvement; p < 0.01); it was significantly higher in the patients with distant metastases than in those without them (p < 0.05). Tumor MMP-9 and MMP-7 concentrations were not significantly associated with the indices of the tumor progression. The patients were followed up for 1 to 85 months (median, 18.3 months). According to the univariate analysis, high (> 32.6 ng/mg protein) MMP-2 and low MMP-7 (< 1.1 ng/mg protein) levels in the gastric cancer tissue represent statistically significant unfavorable prognostic factors for overall survival. Increased TIMP-2 level is associated with a non-significant decrease in the overall survival (p > 0.05), whereas the MMP-9 level was unrelated to the gastric cancer prognosis. Only T index (p = 0.0034) and tumor MMP-7 content (p = 0.026) remained independent prognostic factors in the multivariate regression analysis.Conclusion: The majority of gastric cancer patients demonstrate a significant increase in the expression of three MMP family members, i.e. gelatinases (MMP-2 and 9), and matrilysin (MMP-7), in the tumors, as compared to adjacent histologically unchanged mucosa. Only MMP-2 levels were associated with the disease progression, increasing with higher TNM system indices. High MMP-2 and low MMP-7 content in the gastric cancer tissue are significant unfavorable prognostic factors for the overall survival in the univariate analysis, but only MMP-7 has retained its independent prognostic value in the multivariate assessment.

Rationale: Biochemical diagnostics of neuroendocrine tumors (NETs) is based on the analysis of universal and specific markers, according to the tumor type and clinical manifestations of the disease. Measurement of gastrin levels in the serum of NETs patients is important in the diagnosis of the functioning pancreatic, gastric and duodenal NETs.Aim: To perform a comparative analysis of the results of gastrin measurement as a biochemical marker of NETs using different test-systems.Materials and methods: Serum gastrin was measured in the serum of 30 NET patients and 18 normal controls by immunochemiluminescent (Immulite 2000 Gastrin, “Siemens”) and ELISA (Gastrin-17 ELISA, “Biohit”) assays.Results: The analysis of the results of gastrin measurement by immunochemiluminescent and ELISA methods showed significant variability of its levels, reaching 2016 and 1988 pmol/l, respectively. Comparable analytical sensitivity of the two test systems was confirmed by similar increases (over 30- to 40-fold) of the median levels of the hormone in the serum of patients with gastric NETs, compared to healthy controls. The analysis of diagnostic efficacy accounting for their respective cut-offs demonstrated higher sensitivity (95.4%) of the Immulite 2000 Gastrin test-system, which measures total gastrin, than that of the ELISA method (80.0%), specific to gastrin-17.Conclusion: The analysis shows high efficacy of immonochemiluminescent method of measurement of total gastrin as a biochemical marker of NETs.

Matrix metalloproteinases (MMPs) are a family of zinc-containing endopeptidases that catalyze the degradation reactions of the extracellular matrix components. In humans, 23 enzymes of this family are known, which are subdivided into 6 groups based on their structure and substrate types: collagenases, gelatinases, stromelysins, matrilizines, MMP membrane type and other MMPs. MMP functions are diverse, and an imbalance of their activity may be one of the etiological factors of various diseases. The review considers classification, regulation of activity and genetic polymorphism of matrix metalloproteinases in health and in various pathological processes in the human body. A list of the most studied polymorphic variants of MMP genes is given, their functional effects are described and the results of associative studies are presented.

Background: Despite comparatively favorable clinical course and satisfactory prognosis of endometrial cancer, the increase of its incidence observed in the last years both in this country and abroad requires the development of new approaches to early diagnostics, prognostic assessment and the choice of personalized postoperative management. Therefore, exploration of biological markers for fundamental tumor characteristics, such as invasion, metastasizing, proliferative activity, sensitivity to endogenous and exogenous regulators, are still on the agenda. The family of matrix metalloproteinases (MMPs) degrading the majority of extracellular matrix components and involved in all stages of tumor progression comprise a rich source of such markers.Aim: Comparative evaluation of MMP-2, 7, 9 and their type 1 and 2 tissue inhibitors (TIMP) levels in the serum of patients with endometrial cancer and benign endometrial disease and in healthy women, and the analysis of its associations with the main clinical and pathologic characteristics of cancer for evaluation of their potential diagnostic and prognostic value.Materials and methods: Ninety four patients with endometrial cancer and 53 women with benign endometrial lesions (28 with polyps, 25 with various degree of hyperplasia) were enrolled into the study. The age of cancer patients was 36 to 78 (median 60) years, of those with benign lesions, 20 to 79 (median 54) years. The control group involved 77 practically healthy women aged 19 to 75 (median 46) years. The concentrations of the markers studied were measured by direct enzyme immunoassay (ELISA) kits (Quantikine, R&D Systems, USA).Results: A significant increase of MMP-7 and TIMP-2 levels was demonstrated both in cancer patients (median values 5.5 and 96.9 ng/ml, respectively) and in those with benign endometrial lesions (median values 5.7 and 73.2 ng/ml, respectively) as compared to the control (median values 2.1 and 60.7 ng/ml, respectively). The ROC curve allowed to establish a cutoff level for MMP-7 (3.5 ng/ml) with good sensitivity/specificity ratio for endometrial cancer (88% and 87% respectively); however, it did not allow for differentiating between benign and malignant endometrial lesions. On the contrary, MMP-2, MMP-9 and TIMP-1 levels were 1.2-1.3-fold decreased in patients with cancer and benign endometrial diseases as compared to the control; MMP-9 and TIMP-1 levels in cancer patients were also lower than in those with endometrial polyps and hyperplasia. There were no significant associations between the levels of the markers studied and the indices of progression, histological type and grade of endometrial cancer. Also, no differences between the marker levels in serum of patients with polyps or various endometrial hyperplasias were identified.Conclusion: MMPs and TIMPs assessed in this study cannot be considered as potential diagnostic markers of endometrial cancer; however, they might be useful for disease monitoring, prognosis assessment and evaluation of sensitivity to targeted therapy.

Background: Analysis of free light chains of immunoglobulins (FLC) in the serum is an effective method in the diagnosis of multiple myeloma. Plasma cells produce two types of FLC: κand λ-FLC. FLC, which are not incorporated into monoclonal intact immunoglobulins, are released into circulation, and then are filtered and reabsorbed in kidneys depending on their molecular weight. Circulating FLC commonly form homodimers, known as Bence-Jones protein, which is a biomarker of Bence-Jones multiple myeloma. According to the international guidelines, the ratio κ/λ FLC is an important diagnostic criterion of multiple myeloma. Aim: To evaluate the diagnostic and prognostic value of serum FLC in multiple myeloma patients. Materials and methods: We examined 118 patients with multiple myeloma, admitted to the Department of Hemoblastosis Chemotherapy of N.N. Blokhin Russian Cancer Research Center from 2010 to 2016, and 68 healthy men and women. Serum concentrations of FLC were measured with an immunoturbidimetric method using the test-system Freelite Human Lambda and Freelite Human Kappa (Binding Site Inc.). Results: The levels of monoclonal κor λ-FLC in patients with G-, A-myeloma and Bence-Jones multiple myeloma were significantly higher than those in the control group (p < 0.005). The diagnostic sensitivity of quantification of FLC and their ratio was 87.3% and 89.8%, and in combination with the use of immune electrophoresis it was close to 100%. Analysis of progression free survival and overall survival showed significant differences (p < 0.04) between the groups of patients according their κ/λ FLC ratio. The basal value of κ/λ FLC ratio of less than 0.04 and more than 140 was a predictor of unfavorable outcome. Conclusion: The inclusion of the determination of serum FLC into the assessment plan of patients with suspected monoclonal gammapathy makes it possible to increase diagnostic sensitivity of the available methods for paraprotein determination, as well as to monitor patients with non-secreting multiple myeloma. FLC analysis in multiple myeloma patients acquires special significance in the prognosis of remission, since the antitumor response based on their measurement is seen earlier than that based on the results of standard immunochemistry studies.

Background: The cause of late diagnosis of renal cancer lies in its durable, almost asymptomatic course. Due to the use of antiangiogenic therapies much progress has been made in its treatment in recent years. Yet, many questions concerning the diagnosis, prognosis and prediction of the efficiency of targeted therapy remain unsolved. Therefore, exploration of new renal cancer molecular markers, especially those related to its angiogenic and invasive activities, are still on the agenda. Such markers include the family of matrix metalloproteinases (MMPs) that degrade the majority of extracellular matrix components and are involved at all stages of tumor progression. Aim: Comparative evaluation of MMP2, 7, 8, 9 and type 1 tissue inhibitor (TIMP-1) levels in serum of healthy individuals and patients with renal cancer or benign renal tumors, analysis of their associations with the main clinical and pathologic characteristics of the disease. Materials and methods: We examined 99 renal cancer patients (of those 94 with primary tumor and 5 at progression) and 10 patients with benign renal tumors. The control group included 97 healthy individuals. Levels of the proteins studied were measured using respective direct ELISA kits (Quantikine®, R&D Systems, USA). Results: MMP-7, MMP-8 and TIMP-1 levels in the sera of renal cancer patients were significantly higher than in the control group and in benign renal tumor patients. MMP-2 and MMP-9 levels did not differ significantly between the study and control groups. At MMP-7 cut-off level of 3.0 ng/mL, its diagnostic sensitivity for primary renal cancer was 84%, specificity in relation to “healthy” control – 87.5%, in relation to the pathologic control (healthy donors+benign renal tumor patients) – 73%. The best sensitivity: specificity ratio for TIMP-1 was 67 and 65% at the cut-off level of 315 ng/ml. No cut-off value with acceptable sensitivity: specificity ratio was found for MMP-8. Serum levels of all these 3 markers were positively associated with disease stage and TNM indices; MMP-7 and TIMP-1 levels also increased with lower differentiation grade. In 5 patients evaluated at disease progression the levels of all the markers studied were markedly higher than in the primary patients, and exceeded the estimated cut-off values. Conclusion: MMP-7 should be regarded as the most promising serological renal cancer marker; its serum levels exceed the cut-off value even in 84% stage I patients. TIMP-1 has acceptable sensitivity (70% and above) only from stage II renal cancer onwards, while MMP-8 levels are increased only at stage III–IV of the disease.

Renal cancer (RC) is a common malignancy of the genitourinary system. Clear cell renal cell carcinoma is the most common histological type of RC. In most cases diagnosis and prognosis of clear cell renal cell carcinoma are based on the results of instrumental tests, while search for novel molecular RC markers and their characterization remain relevant. Molecular genetic abnormalities accompanied with changes in gene expression underly the RC carcinogenesis; however, diagnostic panels of the expression markers of RC are still not widely used. This review represents the results of recent research in the area of gene expression markers of RC aimed to elaborate prognostic test systems. Application of the NotI-microarray methodology allowed for identification of many novel genes associated with RC pathogenesis. The relationship of alterations of expression level and methylation of chromosome 3 genes with RC progression and metastasis has been shown. Based on this data, a diagnostic marker system for RC have been proposed with identification of expression and methylation profiles and novel markers, that is an urgent problem in modern urologic oncology.

Background: The progress in cancer treatment, including bone malignancies, is associated with advances in molecular biology. Based on the results of a number of studies, treatment of bone sarcomas have been expanded with targeted therapy that uses drugs with targeted actions, including anti-angiogenic and bevacizumab, in particular. It inhibits the binding of a key activator of neoangiogenesis, vascular endothelial growth factor (VEGF), with its receptors type 1 and 2 (Flt-1 and KDR) on the surface of endothelial cells, which results in a decrease in vascularization and in inhibition of tumor growth. Beyond VEGF, other activators of neoangiogenesis have been identified, such as interleukin 16 (IL-16). Aim: To compare baseline serum IL-16 and VEGF in patients with malignant, borderline and benign bone tumors. Materials and methods: Serum IL-16 and VEGF levels was compared in 138 patients with primary bone tumors: benign (n=10); borderline (giant cell bone, n=22); malignant (n=106), aged 14 to 50 years, by immunoenzyme assay (Biosource, USA for IL-16 and R&D, USA for VEGF) before any specific treatment. Bone malignancies were identified as osteosarcoma (n=45, among them 35 typical, 6 parosteal, and 4 periosteal), chondrosarcoma (n=24), Ewing sarcoma (n=27), and undifferentiated pleomorphic sarcoma (n=7) and chordoma (n=3). Results: The rate of IL-16 identification in the serum of bone tumors patients was 93%, with no significant differences depending on the histological structure of the tumor. No association between the size of primary tumors and IL-16 serum levels was found. Overall 3 and 5-year survival of patients with malignant bone tumors with IL-16 serum levels>33 pg/mL was significantly lower than in those IL-16 levels of≤33 pg/mL. Overall 5-year survival in osteosarcoma patients with higher IL-16 serum levels 1.6-fold lower, in Ewing sarcoma patients, 1.7-fold lower, and in chondrosarcoma patients, 1.8-fold lower than that the patients with IL-16 levels of≤33 pg/mL. VEGF levels in bone sarcomas patients were significantly higher than in those with borderline and benign tumors, whereas statistical analysis did not find any significant difference in VEGF levels depending on the histological structure of the primary tumor. Maximal VEGF levels were found in periosteal osteosarcoma, minimal ones, in parosteal osteosarcoma. Overall 3 and 5-year survival of patients with bone malignancies and serum VEGF concentrations above the mean for the group (> 493 pg/mL) was higher than that in the patients with low VEGF levels. Similar results were obtained in osteosarcoma, whereas in Ewing sarcoma and chondrosarcoma higher 3 and 5-year survival rates were observed in patients with serum VEGF levels below 493 pg/mL. Conclusion: These data suggest that IL-16 and VEGF expression could be associated with pathophysiological changes related to growth and metastatic process of bone sarcomas, and may be a subject for further studies to determine the levels of these biomarkers and their predictive value in bone malignancies.

This review summarizes current data on neuroendocrine tumors (NET), which, unlike other neoplasms, are able to produce biologically active substances (hormones, vasoactive peptides, amines). It is exactly their main characteristic that allows to unify this heterogeneous group and that may determine their clinical course. We present integrated recommendations for biochemical diagnosis and confirmation of over-secretion syndromes based on a panel assessment of NET biochemical markers. Data from the literature are reviewed on evaluation of clinical significance of generic and specific NET markers, as well as the results of the studies performed by the authors themselves. Three hundred and thirty patients were examined with NETs of various localization (pancreas, stomach, small intestine and large intestine, lungs) and with metastatic NET disease with unknown primary location, who were treated in the N.N. Blokhin Russian Cancer Research Center. The control group included 115 healthy individuals. Before and during the treatment, plasma and serum chromogranin A (CgA) and serotonin levels, as well as 5-hydroxyindoleacetic acid (5-HIAA) in a 24-hour urine sample were measured with standardized immunoenzyme plate-based assays (“Chromogranin A ELISA kit”, Dako A/S; “Serotonin ELISA and 5-HIAA ELISA”, IBL International GMBH). We evaluated clinical importance of CgA as a generic NET marker, as well as that of serotonin and its metabolite 5-HIAA as specific markers of the carcinoid syndrome. CgA was shown to be the most efficient biochemical marker for diagnosis, assessment of prevalence and monitoring of NETs. CgA has a high diagnostic sensitivity (63.4 to 88.9%) in various NETs. An association between CgA secretion and prevalence and biological activity of the tumor was confirmed. CgA measurement is particularly important in functionally inactive tumors, where serotonin and 5-HIAA have lower sensitivity, being specific markers of the carcinoid syndrome.

Background: The insulin-like growth factor (IGF) signaling system plays a major role in development and progression of various malignancies including ovarian cancer, and its components are considered as potential diagnostic and prognostic markers and the objects of molecular targeted therapy.Aim: Comparative evaluation of IGF-I and IGF-II, and IGF-binding proteins (IGFBP)-1, 2 and 3 content in blood serum and tumors of ovarian tumor patients, analysis of their associations with key clinical pathologic characteristics of ovarian cancer and evaluation of clinical value of these markers for disease diagnostics and prognosis.Materials and methods: IGF-I, II, IGFBP-1, 2 and 3 levels were measured with standard ELISA kits (Mediagnost, Germany) in blood serum and tumor extracts of 74 patients with ovarian cancer, 16 patients with benign and 14 with borderline ovarian tumors. The control group comprised 77 healthy women. Results: Three potential serological markers of ovarian cancer, namely IGF-I, IGFBP-1, and IGFBP-2 were revealed. The best sensitivity to specificity ratio was demonstrated for IGFBP-2: at a 370 ng/ml cut-off level, these indices were 87 and 79%, respectively. The diagnostic markers found in our study were also associated with the prognosis of overall survival in ovarian cancer. In the multivariate analysis, low IGF-I serum levels retained its independent unfavorable prognostic value. The disease prognosis is influenced also by IGF-II and IGFBP-1 content in the tumor tissue.Conclusion: In ovarian cancer patients, there is a disbalance of IGFs/IGFBPs. Some components of this system could be potentially used as diagnostic and prognostic markers of the disease.

Background: Bone sarcomas are extremely malignant prone to rapid hematogenic metastasing. Evaluation of biological marker expression by the tumor is important not only for the search of new potential chemotherapy targets, but for the assessment of the disease prognosis.Aim: A comparative evaluation of matrix metalloproteinases (MMP)-2, -7, -9 and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the serum of patients with primary bone tumors and in healthy people to identify their potential association with the histological characteristics of the tumor and the disease prognosis.Materials and methods: A comparative study of serum MMP-2, -7, -9, and TIMP-1 levels was performed in 54 patients with primary bone tumors (malignant, 45 patients, including central osteosarcoma in 21, periosteal osteosarcoma in 4, Ewing's sarcoma in 11, primary chondrosarcoma in 6, undifferentiated pleomorphic sarcoma in 3, and borderline giant cell tumors in 9) and in 26 healthy individuals with the use of the immunoenzyme technique (Biosource, USA, for TIMP-1 and R&D, USA, for MMP-2, -7, and -9). Results: The TIMP-1 levels in the serum of patients with central and periosteal osteosarcomas were significantly higher than in the serum of healthy controls (р = 0.038 and p = 0.007, respectively). The MMP-9 levels in patients with bone malignancies were significantly lower than that in the normal controls (p < 0.05). There was a positive correlation between serum TIMP-1 and MMP-9 levels in patients with central, periosteal and Ewing's sarcomas (r = 0.37, p = 0.024). No significant differences in the 5-year survival rates related to serum TIMP-1, MMP-2, -7, -9 levels were found in patients with bone sarcomas. However, in those with osteosarcoma and serum MMP-2 > 160 ng/ml, the overall 5-year survival rate was 1.6-fold higher than in those with lower MMP-2 levels, and in those with ММP-9 levels < 377 ng/ml, the 5-year survival rate was 1.4-fold higher than in patients with ММP- 9 > 377 ng/ml. The worst 5-year survival (33%) was found in the patients with serum ММP-2 levels of less than 160 ng/ml and ММP-9 of more than 377 ng/ml. Conclusion: The results obtain suggest that the expression of MMP-2, MMP-9 and TIMP-1 could be associated with pathophysiological changes related to growth and metastatic process of bone sarcomas and osteosarcoma, in particular. This area could be an object for further studies on levels of these biomarkers and their prognostic value in bone malignancies.

Aim: To develop a highly sensitive real-time polymerase chain reaction (PCR) system for detection of somatic mutations in 542 and 545 codons of exon 9 and 1047 codon of exon 20 of PIK3CA gene comprising more than 80% of all somatic mutations in this gene for application on histological material without macroand microdissection, and to analyze associations between these mutations and clinical and pathological characteristics of breast tumors.Materials and methods: The Allele-specific real-time PCR method with signal detection by TaqMan probes was used. For determination of its analytical sensitivity, plasmids carrying the mutations studied were constructed by standard genetic engineering methods using mutagenesis. DNA samples carrying various mutated/wild type DNA ratios (5.0; 2.0; 1.0; 0.5, 0.25%) were prepared. Results: Multiplex allele-specific real-time PCR method for detection of most common mutations in PIK3CA gene: p.E542K c.1624G>A, p.E545K c.1633G>A, p.H1047R c.3140A>G, p.H1047L c.3140A>T – was developed and optimized.Analytical sensitivity of mutation detection comprised 0.5% for p.E542K and 0.25% for p.E545K, p.H1047R and H1047L enzyme.Conclusion: The method developed for detection of somatic mutations in PIK3CA gene is sufficiently sensitive, specific and efficient, that allows to propose it for a routine screening in clinical diagnostic laboratories for evaluation of disease prognosis and monitoring of response to therapy and its modification.

The review describes the current views on urothelial carcinoma carcinogenesis. Based on the up-todate genetic studies, molecular pathways determining the development of superficial and invasive bladder cancers are considered. It was demonstrated that the development of noninvasive bladder tumors occurred predominantly through oncogenes activation, while the genesis of invasive cancer is based on inactivation of tumor suppressor genes. Principal mechanisms of multiple and recurrent bladder tumors development are discussed including its monoclonal and oligoclonal models.One of the most promising nowadays approaches is the determination of new generation markers, such as molecular genetic abnormalities in the hereditary apparatus of the cell underlying its malignant transformation. Molecular markers' panels used for diagnostics, prognosis, and monitoring of urothelial carcinoma patients are analyzed.

RANK/RANKL/OPG ligand-receptor system is a key player in bone homeostasis regulation directly regulating osteoclast differentiation and osteolysis. Disbalance of bone homeostasis associated with malfunctioning of RANK/RANKL/OPG system underlies such oncological processes as the destruction of bone, metastasis development, tumor progression. Involvement of RANK/RANKL/OPG system in the development of metastasis from various tumors is practically confirmed, but its influence on the development and progression of primary bone tumors still needs thorough evaluation. , In this paper experimental and clinical-laboratory data on the role of RANK/RANKL/OPG system in primary bone tumors pathogenesis available in modern literature are summarized with special attention,paid to giant-cell bone tumor (GCBT) that is already treated with RANK/RANKL interaction inhibitor denosumab. Results of authors study of the levels of RANK/RANKL/OPG system,s components in blood serum of 101 malignant bone tumor (37 – osteosarcoma, 41 – chondrosarcoma, 12 – chordoma, 7 – Ewing sarcoma, 2 – pleomorhic undifferentiated sarcoma, 2 – fibrosarcoma), 32 borderline GCBT, and 30 benign bone tumor patients are also presented. The disturbances in the balance of osteolysis activators and inhibitors in patients with primary bone tumor depending both on the character of neoplasm (malignant, borderline or benign), and histological structure of malignant tumors were demonstrated. The most striking changes in RANK/ RANKL/OPG system manifesting itself in an increase of serum concentrations of all its components and strengthening of association between the levels of soluble receptor and its natural inhibitor OPG were revealed in giant-cell bone tumor patients.The study of the role of RANK/RANKL/OPG system in primary bone neoplasms is a topical goal for clinical investigations; it is also a promising tool for development of new diagnostic methods and for targeted application of specific drugs inhibiting its activity.