EFFECT OF ACTIVATED OXYGEN SPECIES IN HUMAN-LYMPHOCYTES

The cytogenetic effectiveness of activated oxygen species (AOS) generated by the superoxide forming xanthine-xanthine oxidase (X/XO) system was studied in human lymphocyte cultures. The observed chromosome damage was exclusively of the chromatid type. In the experiments a clear dependence of aberration induction on XO concentration and exposure time could be demonstrated. While using anti-AOS agents, the H2O2 antagonist catalase and the hydroxyl radical scavenger formate reduced X/XO induced chromosome damage whereas superoxide dismutase (SOD) did not. In the presence of SOD, aberration frequency was even enhanced. The results indicate that the chromosome damage is caused indirectly via H2O2 formation from spontaneous dismutation of superoxide, whereas H2O2 might be reduced intracellularly giving rise to the highly reactive hydroxyl radical. This effect might be enhanced by SOD, possibly by raising the intracellular amount of easily membrane passing H2O2. Thus, referring to chromosome aberrations, SOD, which is generally reported to protect from AOS, is capable of increasing oxygen mediated biological damage. This observation might be explained by the involvement of DNA associated transition metal, like iron or copper ions, in reducing H2O2. DNA bound copper ions, thought to be necessary for maintenance of DNA quaternary structure, might represent a generator complex for the hydroxyl radical by reduction of X/XO derived hydrogen peroxide. This might cause 'site specific damage' to the DNA which is subsequently converted into chromatid-type aberration by S-dependent misreplication and/or misrepair. This is different to the formation of radiation induced chromosome aberrations which arise by an S-phase independent mechanism.