The Influence of Nimotuzumab in Combination with EGF on the Cell Cycle and Apoptotic Level of Tumor Cells
L. V. Garmanchuk *
Taras Shevchenko National University of Kyiv, Educational and Scientific Centre, “Institute of Biology”, Kyiv, Ukraine
O. I. Dzhus
Taras Shevchenko National University of Kyiv, Educational and Scientific Centre, “Institute of Biology”, Kyiv, Ukraine
V. V. Nikulina
Taras Shevchenko National University of Kyiv, Educational and Scientific Centre, “Institute of Biology”, Kyiv, Ukraine
T. V. Nikolaienko
Taras Shevchenko National University of Kyiv, Educational and Scientific Centre, “Institute of Biology”, Kyiv, Ukraine
O. V. Skachkova
National Cancer Institute, Kyiv, Ukraine
D. V. Shelest
Taras Shevchenko National University of Kyiv, Educational and Scientific Centre, “Institute of Biology”, Kyiv, Ukraine
*Author to whom correspondence should be addressed.
Abstract
Aim: The aim of study was to show the combined influence of nimotuzumab (NTM) and EGF on the proliferation, apoptosis by breast cancer cells line MCF-7.
Methodology: Cell line was cultivated in DMEM under standart conditions. Nimotuzumab and EGF was added to the cells which reached 70-80% monolayer. Flow cytometry assay was conducted to determine cell division in the phases of cell cycle and apoptosis. Cell viability was assessed by MTT test and routine calculation cells used Gor’yaev chamber test with trypan blue.
Results: Obtained results show that the efficiency of the combined effects on tumor cells of breast cancer MCF-7 antibodies to epidermal growth factor receptor – nimotuzumab, in combination with epidermal growth factor enhances the cytotoxic/cytostatic effect of NTM. According to the data, NTM appeared a cytostatic, cytotoxic and proapoptotic effects. Thus, after NTM addition to cultured cells, relative quantity of cells in phases G0 / G1 were increased in 1.2 times (P<0.05), compared to control, the percentage of apoptotic cells was 4-fold increased (P<0.01), and dead cells - almost 2-fold (P<0.05). Previous incubation of cells with EGF (5 nM) during 2 hours before adding NTM was characterized by an increase in the relative content of cells in G0 / G1 phase to 78,3±2,2%, against 73,4±1,4% NTM impact, 58,8±2,1% in the control and 38,9±0,9% for EGF influence.
There was also a redistribution of concentration of apoptotic and dead cells: reducing the percentage of apoptotic compared to the influence of the mono NTM and increase dead cells, respectively.
Keywords: Tumour cells, breast cancer, epidermal growth factor, nimotuzumab