THE BIOLOGICAL PROPERTIES OF TUMOR CELLS IN C57BL/6 MICE WITH TRANSPLANTABLE LEWIS LUNG CARCINOMAWITH INFLUENCE OF ALLOGENEIC MESENCHYMAL STEM CELLS
L. V. Kladnytska1, A. Y. Mazurkevych1, L. V. Garmanchuk2, S. V. Velychko1, V. V. Nikulina2, 4,
T. V. Nikolaienko2, V. V. Kovpak1,O. I. Dasyukevich3, J. V. Skachrova2, A. O. Pavlukova1
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1National University of Life and Environmental Sciences of Ukraine, Heroyiv Oborony str., 15, Kyiv, 03041, Ukraine|
2Taras Shevchenko National University of Kyiv, ESC «Institute of Biology», Ave. Glushkov, 2, Kyiv, 03022, Ukraine
3RE Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, National Academy of Sciences of Ukraine, Vasylkivska str., 45 Kyiv, 03022, Ukraine
4National Institute of Cancer State Institution, Lomonosova str., 33/43, Kyiv, 03022, Ukraine
The purpose of the research — to study the biological properties tumor cells in mice C57BL/6 with transplant Lewis lung carcinoma by the influence of allogeneic mesenchymal stem cells.
Experiments were performed on male mice C57Bl/6 2–3 months of age, weighing 20–22 g. Allogeneic mesenchymal stem cells were obtained by cultivation of primary material that was isolated from the bone marrow of mice C57Bl/6. Cell culture was cultivated at 37 °C, 100 % humidity and 5 % CO2 in DMEM with addition of 20 % fetal bovine serum and 1 % antibiotic-antimycotic. Mice were intramuscularly inoculated cell suspension metastatic Lewis lung carcinoma (LLC) in concentrations 1x106/0.1 ml Hanks solution. After tumor cell inoculation mice were divided into control and experimental groups of 8 animals each. It were injected intravenously allogeneic MSCs 4-th passage in the number 1.25x104 to experimental group mice on day 8 after tumor cells inoculation. On the 20-th day from the beginning of the experiment determined the level of aneuploidy and primary tumor cell division according to the cell cycle phases by flow cytometry. It was established that the use of allogeneic MSCs increased aneuploid cells quantity in 1.3 times in primary tumors and in experimental group mice it was 76.68+1.99*** at p<0.001, whereas in mice not injected MSCs — 59.20±0.80 %. Effect of MSCs on the increase in tumor cell aneuploidy confirms by analysis of variance — ɳ2х=0.86 at p<0.001.
Under the influence of allogeneic MSCs we so changing distribution of tumor cells in cell cycle phases. In particular, the influence of allogeneic MSCs confirmed probable increase the number of aneuploidy pool proliferative cells (S+G2/M) of the primary tumor to 68.02±3.42* against this indicator control group animals 56.17±2.90 % (p<0.05).
The apoptosis level of tumor cells under the influence of MSCs significantly reduced to 41.82±1.37 % (p<0.001), compared with 56.59±0.92 % in the control group animals, which was confirmed by analysis of variance — ɳ2х=0.88 at p<0.001.
Keywords: ALLOGENEIC MESENCHYMAL STEM CELLS, CELL CYCLE PHASES, ANEUPLOIDIYA, FLOW CITOMETRY, LEWIS LUNG CARCINOMA
