Assessment of Cytotoxic Effect Mechanisms of Gas-Discharge Plasma Radiation

The aim of the investigation was to assess the mechanisms of cytotoxic effect of gas-discharge plasma radiation on lymphosarcoma and breast cancer cells.

Materials and Methods. The experiment was carried out on the strains of rat lymphosarcoma (LSR) and breast cancer (RMK1) cells. 4 ml of cell suspension at (4–6)·10⁶/ml concentration was exposed to gas-discharge plasma radiation in various time modes. Plasma radiation was generated by impulse device with the following set characteristics: burst time — 100 µs, voltage — 11 kV, energy per pulse — 5.9·10^-2 J, pulse frequency — 10 Hz. Cytotoxic effect of gas-discharge radiation was assessed using fluorescent dye Hoechst (Sigma ALDRICH, USA), Propidium iodide (Sigma ALDRICH, USA) and МТТ-test. Structural changes in cells were studied by electron microscopy. Cytoplasmic membrane condition was assessed by microviscosity change using a hydrophobic fluorescent probe pyrene (Sigma ALDRICH, USA). The level of oxidative processes was determined by fluorescence of bityrosine, tryptophan, glycated proteins and lipid peroxidation processes. The state of coenzymes was estimated by NAD(P)⁺/NAD(P)Н+Н⁺ and FAD⁺/FADН₂. DNA cell damage degree was assessed by DNA-comet assay.

Results. 600-second radiation exposed to LSR and RMK1 cells was found to be a half-lethal dose. Such radiation causes significant changes in the structure of cytoplasmic and nuclear membranes, intracellular content, reduces microviscosity indices both in a lipid bilayer and in protein-lipid interaction area of LSR and RMK1 cells. Protein molecules of these cells undergo marked oxidative modification exposed to gas-discharge plasma radiation. No accumulation of lipid peroxidation products was recorded. The content of reduced NAD(P)Н+Н⁺ and oxidized FAD⁺ increases in the cells under study under plasma radiation. The number of cells with significantly damaged DNA increases up to 81% by 600^th second of exposure. All changes were in direct relationship to time exposure duration.

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Ivanova I.P., Trofimova S.V., Vedunova М.V., Zhabereva А.S., Bugrova M.L., Piskaryov I.M., Karpel Vel Leitner N. Assessment of Cytotoxic Effect Mechanisms of Gas-Discharge Plasma Radiation. Sovremennye tehnologii v medicine 2014; 6(1): 14