SUMMARY. Heterogenity of the tumor population, the presence of tumor stem cells in it, is one of the reasons for the resistance of tumors to antitumor therapy, recurrence and metastasis, as well as the complexity of treatment of the cancer. The aim of the study was to enrich of the breast adenocarcinoma cells, MCF-7, multicellular tumor spheroids (MTS) with cancer stem cells, enriched MTS (eMTS). Other aim was to investigate the resulting subpopulation of CSC in MTS by biochemical, immunological and cytological methods. According to the results of the study, it was found that in the conditions of lack of nutrient medium, with the addition of certain growth factors, the percentage of CSC in the cell population of MTS increased significantly. The obtained results demonstrated increasing of the CSC subpopulation. It was indicated according to biochemical, cytological and immunological methods simultaneously. Thus, the percentage of CD133+ cells increased from 12.47 to 82.08 %, Nestin+ from 31.3 to 82.58 %. According to immunohistochemical staining data the expression of
other markers of CSC: CD44, CD133, bmi1, also increased. The activity of aldehyde dehydrogenase in MCF-7 cells in monolayer culture was 0.07 mol/mg protein per minute and increased to 1.58 mol/mg of protein per minute in eMTS. The activity of glucose-6-phosphate dehydrogenase (G6FDG) in MCF-7 under conditions of monolayer growth was 934.6 ± 148.3 × 10–6 mol/mg of protein per minute. At the same time in the enriched by CSC MTS the activity of G6FDG increased more than in 1.5 times. The activity of the lactate dehydrogenase (LDH) in MCF-7 cells in monolayer culture was 65.12 ± 1.28 µmol/mg of protein per minute, and in eMTS, decreased in 5.5 times. Thus, based on the analysis of the obtained data, we can assume that under conditions of enrichment of the tumor population by CSC the receptor and energy profile of MCF-7 cells changed. So, MTS are approaching to characteristics of metastatic micronode, and tumor cells are approaching to cancer stem cells.
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