Diagnostic biomarkers cancer diagnostics at preclinical stage seem to be a very promising strategy to increase effectiveness of anti-cancer treatment. Currently there are no such biomarkers available for daily routine practice. However, there are some candidate molecules in research that possibly can be used as biomarkers for early diagnosis, one of them is microRNA. MicroRNA is a small, 20–25 bp, non-coding RNA that is highly involved into epigenetic regulation of gene expression. These molecules participate in malignant transformation of normal cells into cancer cells including melanoma. And moreover, definite expression level of some microRNAs are essential for normal differentiation and function of human cells. Changes in microRNA profile are one of the reasons for malignant tumor development. Identification of these changes may help to develop diagnostic systems to start anti-cancer treatment at early stages.

According to modern data, metformin is a unique drug, which can act not only as sugar-reducing medicine, but also as an antiproliferative element. More and more new researches about metformin effects in oncologic patients appear during last decades. A lot of researchers suppose that metformin is a new promising medicine for chemopreventive and neoadjuvant cancer therapy. This literature review covers current researches of metformin in context of it»s possible antiproliferative effects.

The purpose of this study is to analyze methods of mathematical modeling for calculating the stage of primary sublimation, as the most important stage in lyophilization technology. Presented are mathematical formulas, equations for the calculation of heat and mass transfer processes, during the removal of 90 % of all frozen ice. A model is considered that takes into account the contribution of all thermal effects, including the transient energy balance, taking into account the heat transfer through the side wall of the vial and radiation, even if they are present in a small amount. The mathematical model can be used to optimize the lyophilization cycle, and also as tools for technological monitoring (using sensors based on models). The model considered in the article is a one-dimensional nonstationary state model in which the correct comprehensive transient energy balance has been introduced to describe the heat transfer through the glass of the vial, and the results are estimated using experimental data. The equations used in the simulation describe the mass and energy balances in the dried layer, taking into account the rate of adsorption/desorption of water at the interface, mass and heat transfer at the sublimation interface, as well as the energy balance of heat transfer in the wall of vials, shelf and other factors affecting the process of sublimation. Conclusions are made on the presented mathematical models and the characteristic of the direction of the process of optimization of primary sublimation in lyophilization technology is given.

Introduction.Autophagy, a catabolic process of protein and organelle recycling by transferring defective cytoplasm and organelles into double-membraned vesicles to degrade and regenerate materials, plays a critical role in maintaining energy homeostasis. Inefficiency chemo-and radiotherapy is largely associated with the activation of autophagy. Among the metals needed by the living organism, iron occupies a special place. The rapid growth of malignant tumors requires much more iron than the metabolism of normal cells.

Objective.To elucidate the relationship between autophagy and iron in melanoma progression.

Materials and methods.In this study we used 2D- and 3D-culturing of melanoma cells with high expression of CD71 (mel P and mel Z) and low expression of CD71 (mel Gus and mel Ibr), flow cytometry and fluorescence microscopy.

Results.The uptake of iron in cancer cells occurs through translocation of the complex of transferrin/receptor (CD71) in the cytoplasm with subsequent dissociation of iron from the complex. Chelation of iron by deferroxamine in melanoma cells mel P and mel Z reduced the level of autophagy about 2-fold. In the presence of an iron donor ferrum ammonium citrate the level of autophagy increased 2.5- fold. The same correlation was observed in melanoma cells with low expression of CD71. Chelation of iron in melanoma cells with high CD71 expression blocked the formation of capillary-like structures. In the presence of an iron donor the formation of capillary-like structures was also not observed. The same correlation was observed in melanoma cells with low expression of CD71. There was an increase in CD105 expression about 50 ± 5 % and 800 ± 50 % under the condition of iron chelation in melanoma cells with high and low expression of CD71, respectively. Quite unexpectably, iron donor also increased expression of CD105 about 35 ± 4 % and 300 ± 3 % in melanoma cells with high and low expression of CD71, respectively

Conclusions.The activation of autophagy promotes the survival of tumor cells by triggering a number of metabolic functions with the participation of iron.

Intrоduction.Human antigen PRAME is preferentially expressed in a number of different tumor types and may be a potent target for anti-tumor immunotherapy.

Purpose.To study anti-tumor action of immunogenic mix recombinant PRAME protein and adjuvant in mice with innate immunity.

Materials and methods.C57BL/6 female mice were used for immunization with purified human recombinant protein PRAME. Human PRAME gene coding sequence was cloned in mammalian expressing vector pCEP4 and resulting plasmid was introduced in mouse melanoma B16F10 cells by transfection followed by RQ-PCR, Western blot and flow-cytometry analysis. Then stably PRAME-transfected melanoma cells were injected in mice.

Results.The mouse melanoma B16F10 cells stably expressing human PRAME protein were obtained. We demonstrate the 10-fold decreased tumor volume in mice with melanoma B16F10 expressing human PRAME after preventive immunization series with recombinant PRAME protein. The tumor volume reducing was correlated with high titer (6.14 × 10 5) of anti-PRAME antibodies in mice sera.

Conclusion.These data indicate that recombinant protein PRAME is immunogenic and may be a potent antigen for immunotherapuetics studies.

Introduction.Phyto-anti-estrogen secoisolariciresinol (SECO) has similar effectiveness to that of Tamoxifen (TAM), a member of selection estrogen receptor modulators, in estrogen-positive models of murine mammary adenocarcinoma and human breast cancer in vivo. Due to its antagonistic and agonistic functions Tamoxifen may enhance risk of development of uterine adenocarcinoma while providing effective prophylactics of breast cancer metastases. SECO effect on proliferative activity of circulating or disseminating tumor cells (occult metastases) is still unclear. We used epithelial cell marker – intracellular cytokeratin 19 (CK19) to study SECO function in terms of possible metastatic process, since prognostic significance of CK19 is well established for identifying occult metastases and breast cancer dissemination.

Objective.To evaluate risk of tumor cell dissemination in mice with transplanted mammary adenocarcinoma after exposure to SECO and TAM.

Materials and methods.Mice BDF 1 [C57Bl6j × DBA2] bearing Са755 of the 3rd passage were used for the experiments. CK19 expression was evaluated 24 hours after 10-day course of SECO in the effective single doses of 250 mg/kg or ТАM 50 mg/kg. Flow cytometry, immunofluorescence with light and luminescence microscopy were performed to evaluate CK19 expression.

Results.Parameter GMFI ± SD (geometric mean fluorescence intensity ± SD) for CK19 expression in SECO and TAM groups in blood accounted for 28.87 ± 13.70 and 28.02 ± 9.50 and in control tumor growth (CTG) group GMFI ± SD was 31.94 ± 5.02; while in bone marrow it was 30.14 ± 2.33, 39.07 ± 2.30 and 32.48 ± 3.75, respectively.

Conclusion.The results of the study showed similar expression of epithelial intracellular marker CK19 in blood in the studied groups of mice bearing mammary adenocarcinoma Ca755 sensitive to SECO and TAM exposure. GMFI for CK19 expression in bone marrow was lower in SECO group than in TAM (р = 0.0003). The data obtained in the murine model demonstrated no enhanced risk of tumor cell dissemination while performing treatment by phyto-anti-estrogen SECO in therapeutic regimen.

Objective.Determine the frequency of occurrence of tyrosine kinase expression of the mutated ALK and TAG-72 gene among patients with primary melanoma of the skin, to identify their association with a number of histological parameters, and to assess the diagnostic value of the determination of ALK and TAG-72.

Materials and methods.Paraffin blocks with surgical material from 40 patients with primary skin melanoma. For routine histological examination, the material was fixed with 10 % neutral formalin for 24 h, poured into paraffin, sections were prepared with a thickness of 4–5 μm, stained with hematoxylin and eosin. IHC study with monoclonal antibodies D57.3 to ALK was performed on an immunostender – Ventana, with antibodies B72.3 to TAG-72 – on Thermo Fischer. As a detection system used: Envision – for TAG-72 and Ventana – for ALK.

Results.ALK mutation was detected in 7 (12 %), TAG-72 – 4 (10 %) cases. Evaluation of the correlation force between the presence of ALK and TAG-72 showed a direct average coupling strength (correlation coefficient was 0.31). A direct correlation of the mean force between the presence of TAG-72 oncoprotein, ALK mutation and ulceration in the patient was found – the correlation coefficient was 0.53 and 0.68, respectively. There was a statistically significant association between the presence of ALK and lymphoid infiltration, which in most cases (57 %) was pronounced (p <0.05).

Conclusion.Comparing the positive sign of the expression of ALK – 17.5 % and TAG-72 – 10 %, and the positive of their simultaneous detection – 7.5 %, it can be concluded that further studies to determine their diagnostic value are promising. The presence of severe lymphoid infiltration in ALK-positive patients claims a diagnostic value in primary skin melanoma.

Introduction.Bacteriochlorins are the most promising photosensitizers absorbing in the near-infrared spectral region. Their use can enhance the efficiency of photodynamic therapy due to the deeper penetration of radiation into the tumor.

Objective to conduct a preclinical study of the photoinduced antitumor activity and biodistribution of Bacteriosens.

Materials and methods.Bacteriosens is a preparation based on meso-tetra(3-pyridyl)bacteriochlorin absorbing at 747 nm. Photoinduced cytotoxicity was investigated in vitro using human tumor cells: A549, Hep 2, BT-474, MCF-7, SK-BR-3, PC3, and EJ and murine tumor cells: S37, C26, and LLC. In vivo studies were performed in mice with large and small tumors (S37, LLC, and C26).

Results.In vitro investigation show that bacteriosens during optical irradiation led to the effective suppression of tumor cell growth in culture (the IC50 value varied from 0,08μМ to 1,21 μМ) and had no toxicity without exposure to light. The effective photodynamic therapy regimen using Bacteriosens in mice with inoculated small and large tumors of different genesis resulted in regression of a primary tumor node on 90–100 % of the animals in the absence of tumor recurrence within 90 days after treatment.

Conclusion.Bacteriosens is a promising agent for the photodynamic therapy of small and large tumors; it can be successfully used as an alternative, organ-sparing minimally invasive treatment for malignant tumors, including prostate cancer.

Introduction.The development of high-quality domestic reproduced dosage form of сisplatin is necessary to improve the treatment conditions of cancer patients.

Purpose of research.Comparative preclinical study of acute toxicity of newly developed in SIC Oncology reproduced dosage form Cisplatin-RONC® with commercial preparation Cisplatin-Teva.

Materials and methods.The standard methods of estimation of acute toxicity of preparations on small laboratory rodents are used.

Results.Acute toxicity of the compared forms of cisplatin is not significantly different from the effect on the survival and body weight of experimental animals.

Summary.Compared generic and commercial formulations cisplatin almost equitoxic after a single intravenous administration to mice and rats.

Introduction.This article presents a fragment of a preclinical toxicological study of a new Russian anticancer drug derived from n-glycoside indolokarbazole LCS-1208 – study of cardiotoxicity, which is one of the specific complications of anticancer chemotherapy.

Objective.Preclinical toxicological study of the effect of the drug LCS-1208 on the cardiovascular system of animals to assess its cardiotoxic effects.

Materials and methods.Studies were conducted on 40 healthy non-harmless mongrel male rats and 4 dogs Beagle, male and female. The drug was administered daily 15 times to rats-intraperitoneal in total doses of 50, 100 and 200 mg/kg; to dogs – intravenously in total doses of 20 and 30 mg/kg. The period of observation of rats was 30 days, for dogs was 60 days. Changes in electrocardiogram indices, macroscopic and histological picture of heart changes and changes in biochemical parameters of enzymes activity – lactate dehydrogenase and aspartate aminotransferase were evaluated.

Results.In rats LCS-1208 throughout the period of observation caused functional changes in electrocardiogram: increase in PQ and QT intervals and cardiac rhythm disturbance (loss of R wave), which indicates a violation of electrical conductivity. Morphological changes in the heart muscle were detected on the 3rd day of observation in total doses of 100 and 200 mg/kg, which remained until 30 days of observation only in animals receiving the drug in the total dose of 200 mg/kg. In some dogs for different periods of observation the drug caused functional changes in the electrical activity of the heart: an increase in the QRS interval, the inversion of the T wave, the appearance of a deep Q wave and an increase in the activity of lactate dehydrogenase and aspartate aminotransferase compared to back ground indicators. Morphological changes in the heart muscle were detected on the 3rd day of observation only in the total dose of 30 mg/kg, which persisted up to 60 days of observation.

Conclusion.It was found that the new Russian anticancer drug LHC-1208, a derivative of indocarbazole N-glycoside, has a cardiotoxic effect, causing functional changes in the cardiovascular system of rats in all doses studied, and in dogs only in the maximum dose. Morphologically, cardiotoxicity is not confirmed in animals receiving a minimal dose of the drug, but only in animals receiving the maximum dose of the drug.

Introduction.New antitumor multitarget drug LCTA-2034, obtained in Gause Institute of New Antibiotics, has demonstrated high activity against prognostically significant transplantable mice tumors by the oral application.

Objective.To investigate the toxicological properties of LCTA-2034 by the oral route of administration on rats.

Materials and methods.Toxicological study of LCTA-2034 was performed on 30 male Wistar rats. Drug substance dissolved in potable water. 2 % solution was administrated per os at the 1 and 5 therapeutic dose (15 × 20 mg/kg or 15 × 100 mg/kg with 24-h interval). During the study dynamics of body weight, hematological parameters, blood biochemical parameters, electrocardiography and urinalysis were performed for all animals. Five animals in each group were sacrificed 1 and 30 days post treatment. The internal organs were subjected to histological evaluation.

Results.The results of the study demonstrate that the treatment with low dose of LCTA-2034 does not produce any changes in majority of examined clinical-laboratory parameters with the exception of urinalysis revealed hematuria on day 1 post treatment. Microscopic pathology observation showed structure abnormalities of varying severity in liver, kidneys, heart, stomach, jejunum, ileum, spleen and thymus. Administration of high dose of LCTA-2034 caused mortality of 2 rats in group. The rest of the rats were observed a body weight lag, decrease of total leukocyte and erythrocyte count, hemoglobin and hematocrit level, relative weight of the thymus. Erythrocytes and nitrates were found in urine both on day 1 and on day 30 post treatment. In groups treated with high dose of the drug in addition to the listed above organs damage of the structure of lymph nodes, pancreas, ileum and brain was detected. Conclusion. Revealed toxic properties of LCTA-2034 depended on dose. Multiple administration of 1 therapeutic dose of the drug produces transient toxic effects completely reversible within 30 days.