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Information × Registration Number 0219U000712, 0116U000039 , R & D reports Title Development of cell test system to evaluate the efficacy and mechanisms of action of antineoplastic drugs. popup.stage_title Head Sydorenko Myhail V., Registration Date 19-02-2019 Organization "Department of Biotechnical Problems in Diagnostic of Institute of Problems of Cryobiology and Cryomedicine National Academy of Sciences of Ukraine" popup.description2 Abstract. Research report: p. 50, fig. - 19, sources of references -95. During the reporting period, using the established test system and a system of indicators, screening was conducted of the effect of classical anticancer drugs and the latest compounds on the biochemical and cytological indicators of monolayer and spheroid cultures. Research object: MCF-7 tumor spheroids, Doxorubicin, Methotrexate, Cisplatin. Cytological, molecular biological methods, biochemical and statistical methods of analysis were involved in the work. The aim of the study was to compare the reactivity of monolayer and spheroid cultures under conditions of modification of microenvironment factors. Determination of proliferative, morphological, kinetic characteristics of spheroids and a monolayer culture under conditions of incubation with anticancer drugs. The objectives of the study were to determine differences in the sensitivity of tumor cells in monolayer and spheroidal cultures and approbation of the cell test system on the latest antitumor compounds. As a result, it was found that tumor cells in a 2D culture are more sensitive to the effects of drugs than in a 3D culture. At the same time, the difference in the percentage of living cells is statistically significant with increasing concentration of the drug. So, with a Doxorubicin concentration of 0.1 ?g/ml 1.0 and 10.0 ?g/ml and an incubation time of 144 hours, the percentage of living cells in a 2D culture was 58.0%, 47.2% and 41.0%, respectively, in 3D - 78.0%, 81.0% and 100%, relative to the control. A similar trend has been demonstrated when cells are cultured with cisplatin. So in a 2D culture, incubation of 144 hours with cisplatin at concentrations of 0.01 ?g/ml, 0.1 ?g/ml and 1.0 ?g /ml resulted in a decrease in the survival rate of cells in a monolayer culture to 73.5%, 48.2% and 17.1%, relative to control. In a spheroid culture, survival rates are significantly higher, namely, 114.0%, 64.5% and 29.6% at appropriate concentrations. When cells were incubated with methotrexate, cell resistance to this preparation was observed in both monolayer and spheroid cultures. But with a methotrexate concentration of 0.1 ?g/ml - 1.0 and 10.0 ?g/ml and an incubation period of 144 hours, the percentage of living cells in the 2D culture was 58.8%, 41.2% and 29.0%, respectively, in 3D - 71.0%, 64.8% and 54.9%, relative to the control. Therefore, as a result of the work, it was concluded that monolayer and spheroid cultures equally reflect the correlation trends of sensitivity / resistance of tumor cells to the concentration of anticancer drugs (cisplatin, doxorubicin, methotrexate), resistance of tumor cells MCF-7 to methotrexate was detected both on monolayer and spheroid culture. At the same time, dose-dependent sensitivity to cisplatin and doxorubicin was indicated in both cell models; anticancer drugs (doxorubicin, methotrexate, and cisplatin) have been studied to have a cytotoxic effect on MCF-7 cells in monolayer culture and inhibit the growth of multicellular tumor spheroids; during long-term cultivation, the studied drugs had a significant cytotoxic effect on tumor cells in monolayer culture, while the sensitivity of cells in spheroid culture was statistically significantly less. Scientific and practical significance. The data obtained indicate a lower sensitivity of the cells of the 3D spheroid culture under changes in the microenvironment and under the influence of anticancer drugs. These circumstances confirm the thesis that the testing of anticancer drugs on the model of multicellular tumor spheroids is an important stage of preclinical testing, which allows you to get more complete information about the possibilities of the drug to influence the most aggressive population of tumor cells capable of migration and invasion, the existence in a state of rest and of conditions nutritional deficiencies. Key words: MCF-7 spheroids, Doxorubicin, Methotrexate, Cisplatin. Product Description popup.authors Железна Мілана Володимирівна Лебедєва Тамара Миколаївна Перепелиціна Олена Михайлівна Сидоренко Михайло Васильвич Юрченко Наталя Петрівна Ястребова Олена Вікторівна popup.nrat_date 2020-04-02 Close