Application of EPR spectroscopy to determine the influence of simvastatin concentration on free radicals in A-375 human melanoma malignum cells

摘要

Malignant melanoma is the most aggressive and lethal form of skin cancer. Therefore the search for new methods of the treatment for melanoma is needed. In this work simvastatin was tested as the substance of potential anticancer activity. The aim of this study was to determine the effect of simvastatin in different concentration on free radicals in A-375 human melanoma malignum cells. Free radicals and growth of A-375 melanoma cells cultured without and with simvastatin were examined. Free radicals were tested by the use of electron paramagnetic resonance (EPR) spectroscopy. o-Semiquinone free radicals existed in A-375 cells. Free radicals of the control A-375 cells and the cells cultured with simvastatin were homogeneously broadened. The growth of A-375 cells did not change for concentrations of simvastatin 0.1 mu M and 0.3 mu M, and it slightly decreased for concentration of this substance equal 1 mu M, so these concentrations of simvastatin were not preferred for therapy. For the concentrations of simvastatin higher than 1 mu M the growth of A-375 cells was considerably weakened. Simvastatin in the concentrations of 0.1 mu M and 1 mu M decreased free radical concentrations in A-375 cells. The concentrations of simvastatin of 3 mu M and 5 mu M accompanied by the strong cell damage and strong formation of free radicals in them were recommended for anticancer therapy of A-375 melanoma cells.