Aktivnost superoksid-dismutaze u animalnoj ćelijskoj kulturi CHO-K1 nakon tretmana fulerenolom i mitomicinom C

Abstract

Eukaryotic cell survives in predominantly reduced conditions. Homeostasis of cellular redox system is an imperative of cell surviving and its normal metabolism. ROS are well recognized for playing a dual role as both deleterious and beneficial species, since they can be either harmful or beneficial to living systems. These species are mutagenic compounds known to lead to DNA damage, favor cell transformation, and contribute to the development of a variety of malignant diseases. All the effects of oxidants are influenced by the cellular antioxidant defenses. This multilayer system consists of low molecular weight components and several antioxidant enzymes. Superoxide dismutases (SODs) are the only enzymes dismuting superoxide radicals. Mitomycin C, a cross-linking agent, demonstrated genotoxicity in all in vitro and in vivo test systems in mammalian cells and animals. Water-soluble fullerenes are well known as cytotoxic agents for many cell lines in vitro. At the other side, fullerenols are good free radical scavengers and antioxidants both in vitro and in vivo. This paper investigates the effects of fullerenol on survival and fullerenol/ /mytomicine (MMC) treatment on superoxide-dismutase (SOD) activity in CHO-K1 cells. Samples were treated 3 and 24 h with fullerenol (C60(OH)24) at concentration range 0.01-0.5 mg/mL and survival was monitored with dye exclusion test (DET). The activity of total SOD was estimated in samples treated with chosen concentrations of fullerenol and MMC (0.5 and 0.1 mg/mL) after 3 and 24 h of cell incubation. Increasing of C60(OH)24 concentration leads to decreasing of percent of surviving cells 3 and 24 h after incubation. The activity of total SOD enhanced with higher concentration of fullerenol, while decreased in the highest concentration at both experimental points. In samples treated with MMC, as well as in samples treated with fullerenol (0.0625 mg/mL) + MMC was noticed boost in total SOD activity in comparison with controls. Treatment with fullerenol decreased SOD activity in rest of samples treated with MMC. Decreased activity of superoxide-dismutase in almost all samples treated with fullerenol and MMC might be contributed to antioxidative properties of fullerenol. Increased enzyme level at concentration of 0.0625 mg/mL may be due to its prooxidative activity.

U ovom radu ispitivani su efekti fulerenola (C60(OH)24) na preživljavanje, kao i tretmana fulerenolom i mitomicinom c (MMC) na aktivnost ukupne superoksid-dismutaze u CHO-K1 (ovarijalnih ćelija hrčka) ćelijskoj liniji. U uzorcima ćelija tretiranim fulerenolom koncentracija 0,01-0,5 mg/mL, praćeno je preživljavanje testom odbacivanja boje (DET) u 3 i 24 h tretmanu. Aktivnost superoksid-dismutaze (SOD) merena je u uzorcima tretiranim fulerenolom izabranih koncentracija i mitomicinom c (0,5 i 0,1 mg/mL) nakon 3 i 24 h. Sa porastom koncentracije (C60(OH)24) opada procenat preživelih ćelija tokom 3 i 24 h. Aktivnost SOD raste sa porastom koncentracije fulerenola i u najvećoj koncentraciji opada u obe vremenske tačke eksperimenta. U uzorcima tretiranim fulerenolom i MMC došlo je do smanjenja aktivnosti SOD, izuzev pri koncentraciji fulerenola od 0,0625 mg/mL, kada je zapažen porast aktivnosti SOD u odnosu na kontrolne grupe.