Vol 15, No 3 (2016)

Introduction. E-cadherin aberrant expression or complete loss is common for a number of human malignant neoplasms, and can be a launching mechanism of an epithelial-mesenchymal transition. Passing through epithelial-mesenchymal transition could in turn promote to the acquisition of so called cancer stem cell phenotype by the transformed cells. The objective of the present study is to reveal the influence of E-cadherin expression level on the amount of cancer stem cells in human colon cancer cell line HCT116. Materials and methods. We have created cell sublines with E-cadherin up- and downregulation and assessed the percentage of cancer stem cells using tumor formation assay, clonogenic assay; we also evaluated profile of cell pluripotency markers. Results and conclusion. We have shown that the proportion of cancer stem cells in human colon adenocarcinoma cell line HCT116 depends on the E-cadherin expression level. E-cadherin expression downregulation results in elevated expression of pluripotency genes and in the increase of proportion of cancer stem cells via activation of Wnt/ß-signalling pathway. E-cadherin upregulation has a reverse effect and decreases the amount of HCT116 cancer stem cells. Thus, E-cadherin expression restoration seems prospective in colorectal anticancer therapy.

Objective. The aim of the study was to research effect of new cyclic hydroxamic acids (CHA) I-VI on activity of Ca²+-ATPase of sarcoplasmic reticulum and cyclic guanosine monophosphodiesterase. Materials and methods. Activity of Ca²+-ATPase of sarcoplasmic reticulum and cyclic guanosine monophosphodiesterase has been evaluated. Results. It has been shown that at studied concentration (0.1 mM; 0.01 and 1 mM), CHA I-VI separate hydrolytic and transporting functions of Ca²+-ATPase of sarcoplasmic reticulum. Therefore, the antimetastatic effect of these compounds is assumed. Thus, at concentration of 0.1 mM, CHA inhibit active transport of calcium through the sarcoplasmic reticulum membrane by 40 ± 4 % (CHA-I), 50 ± 5% (CHA-II), 53 ± 5% (CHA-III), 70 ± 8% (CHA-V) and 75 ± 8% (CHA-VI) and inhibit hydrolysis of ATP by 20 ± 2%, 0 %, 0 %, 45 ± 5 % and 47 ± 3 % respectively. CHA-III, CHA-V and CHA-VI inhibit active transport of Са²+ by 46 ± 5 %, 47 ± 5 % and 60 ± 6 %, and not inhibit or inhibit by 23 ± 2 % and 27 ± 3 % respectively, hydrolysis of ATP at concentration of 0.01 mM. CHA-V inhibits reversibly and non-competitively the hydrolytic function of Ca²+-ATPase of sarcoplasmic reticulum with K = 4 x 10^-5 M. All studied CHA inhibit activity of cyclic guanosine monophosphodiesterase at concentration of 0.1 mM, by less than 20 %. Conclusion. The data obtained predicts the antimetastatic activity of compounds CHA-II, CHA-III, CHA-V, CHA-VI. We recommend to study of CHA-II, CHA-III, CHA-V, CHA-VI on animal models as promising antimetastatic drugs.

Background. Aranose is alkylating antineoplastic agent with cytotoxic and cytostatic properties. Liposomal form of aranose has advantages compared with traditional dosage form. Another alkylating agent, cisplatin, promotes mRNA increase expression level of genes, which control external apoptosis way. Objective. We assume that liposomal form of aranose can change the CD95/Fas, TNFR1, TNFR2, TRAIL, DR3 and DR4/5 expression level in melanoma cell lines mel Kor, mel Z, mel Mtp, mel Mtp-X, mel Ibr, mel Ch, mel Is and mel R. Materials and methods. There are two forms of aranose were used in this work: liposomal form and traditional diluted form for injection. Melanoma cell lines were incubated with both forms. We used RQ PCR to evaluate mRNA expression mRNA expression of DR3, DR4/5, CD95/Fas, TNFR1, TNFR2 and TRAIL relatively gene ABL. Results. CD95/FAS mRNA expression level was most changeable. Liposomal aranose increase CD95/FAS expression level in mel Ibr and mel Mtp-X cell lines and reduced CD95/FAS expression level in mel Ch and mel Is. Conclusion. Thus, liposomal aranose can change mRNA CD95/FAS expression level in melanoma cell lines. Using this process, liposomal form can affect the apoptotic cell death of melanoma.

Objective. The aim of the study was to research the toxicity of L-asparaginase Was79 in rabbits. Materials and methods. Chronic toxicity of L-asparaginase Was79, obtained by modification of native enzyme Wolinella succinogenes in Research Institute of Genetics and Selection, was performed in male and female Russian chinchilla rabbits. L-asparaginase was injected intravenously at the 1 and 5 therapeutic dose (15 x 100 IU/kg or 15 x 500 IU/kg with 24-h interval). The following parameters were tested: body mass, clinical and biochemical blood tests, urinalysis, electrocardiography, pathomorphological evaluation of internal organs. Resutls. The results of the study suggest that the treatment with L-asparaginase Was79 does not influence on the function of heart and kidneys, but damages their structure. Loss in body mass, diarrhea and alteration of stomach and intestine mucosa could be interpreted as evidence of gastrointestinal toxicity. Hematological toxicity was exhibited as a decrease of total leukocyte count, lymphocyte and neutrophils count level in peripheral blood and atrophy of lymphoid tissue of the spleen, thymus and lymph nodes. Elevation of total and direct bilirubin in serum and histopathological findings in liver were found in groups treated with both high and low doses of Was79. Conclusion. Most of these abnormalities were reversible and dose-dependent.

Background. In N.N. Blokhin Russian Cancer Research Center has been developed injectable dosage form based on the substance of indolocarbazole derivative LCS-1208, studied its antitumor activity and mechanism of antitumor effect, which was the basis for the pre-clinical study of the drug toxicity. Objective. The study of subchronic toxicity of formulation of glycoside derivative of indolocarbazole - LCS-1208 after intraperitoneal application to rats. Materials and methods. Dosage form of study drug: lyophilisate for preparation of solution for injection of 9,0 mg in vial. The study was conducted on 40 healthy non inbred male rats, which were obtained from the breeding of the N.N. Blokhin Russian Cancer Research Center. It was studied the action of the drug on the peripheral blood, the functional state of the liver, kidney and gastrointestinal tract of animals. Results. The drug LCS-1208 by daily intraperitoneal administration to rats for 15 days in the 3 studied doses (total dose 200 mg/kg, 100 mg/kg and 50 mg/kg) did not cause death of animals, had no effect on the general condition of the animals, did not cause external manifestations of toxicity, did not change behavioral responses of animals and had no effect on the functioning of the studied organs. Conclusion. The results obtained by the study of the subchronic toxicity in rats were the basis for further preclinical toxicology study of the drug LCS-1208.

Objective of the research was determination level of RRM1 expression in ovarian cancer tissue. Materials and methods. To avoid disadvantages of those evaluations, the present study of the RRM1 expression level in ovarian cancer tissue determination was held using strictly quantitative immunofluorescence flow cytometry-associated method developed by the authors. Results. RRM1 expression was detected in 100 % of samples of ovarian serous adenocarcinoma, with significant differences in the indicator level in variable patients. The high level of RRM1 (≥ 40 % of cells expressing the marker) was detected in 65 % of the tumor samples studied, low (< 40 %) - in 35 % of cases. Data on the lower expression of RRM1 in one-third of the test samples from primary serous ovarian cancer corresponds with clinical observations of gemcitabine monotherapy efficiency in 25-30 % of patients. Conclusion. The authors believe that a strictly quantitative determination of the marker level in tumor tissue can help to overcome the ambiguity of RRM1 clinical significance estimation in gemcitabine treatment effectiveness prediction.

Background. Beta-III tubulin (TUBB3) is a tumor-specific isoform of the microtubule protein beta-tubulin. TUBB3 is considered to be a marker of adverse prognosis and tumor resistance to therapy with taxanes and Vinka alkaloids. Association between TUBB3 expression and histological type of the tumor has not been studied properly yet. Objective. The expression level of TUBB3 in non-small cell lung cancer biopsy specimens has been measured on 2 groups of patients with adenocarcinoma and squamous cell carcinoma. Materials and methods. The samples with adenocarcinoma (n = 43) and squamous cell carcinoma (n = 39) were converted to suspension, filtered, fixed with 4 % formaldehyde, stained with monoclonal antibodies for TUBB3 and DyLight 650-conjugated secondary antibodies to mouse IgG and analyzed by flow-cytometry. The method was developed in N.N. Blokhin Russian Cancer Research Center. Results. The average level of TUBB3 expression in the adenocarcinoma group is higher than in the squamous cell carcinoma group (33.1 ± 12.4 % of cells expressing the marker in adenocarcinoma vs. 26.0 ± 13.6 % in squamous cell carcinoma; differences were statistically significant). The average level of TUBB3 expression in adenocarcinoma is 29.7 ± 8.1 % in female and is 34.9 ± 13.9 % in male (differences statistically insignificant). Since the group of patients with adenocarcinoma was presented by both men and women, while the group of patients with squamous cell carcinoma was only men, from the analysis was excluded all female patients. Differences between the groups remained statistically significant. Conclusion. TUBB3 expression in tumor tissue does not depend on the gender, at least among patients with adenocarcinoma of the lung, and at the same time, the level of TUBB3 in adenocarcinoma tissue is higher in comparison with squamous cell carcinoma tissue.