DNA damage by hydroquinone in human white blood cells: analysis by alkaline single-cell gel electrophoresis.
The genotoxicity of hydroquinone (HQ) in human white blood cells was investigated by means of alkaline single-cell gel electrophoresis (SCGE). The exposure of purified lymphocytes to HQ (0.5-50 microg/ml) produced significant and dose-related increases in DNA migration; conversely, no induction of DNA damage was observed in leukocytes after in vitro treatment of whole blood samples (100-500 microg/ml). Similar differences in DNA damage between whole blood samples and purified lymphocytes were observed after treatments with hydrogen peroxide (H2O2, 50 microM). The DNA damaging activity of HQ was significantly (p<0.001, U-test) inhibited by exogenous catalase (250 U/ml), indicating the generation of peroxides in the mechanism of genotoxicity of HQ. Parallel experiments using the standard SCGE protocol, and an acellular method entailing the lysis of cells before HQ treatment, provided fairly similar results, indicating that HQ oxidation does not require endogenous metabolism. Experiments to compare the effectiveness of HQ in the induction of single-strand breaks and alkali-labile sites in resting cells and micronuclei in cytokinesis-blocked cells indicate that despite the extensive DNA damage detected by SCGE immediately after treatment, a significant excess of micronuclei is not observed after stimulation and in vitro cultivation. These data explain the apparent discrepancy between the high DNA damaging potential of HQ in human lymphocytes, as revealed by SCGE, and the relatively low activity reported in most cytogenetic assays with HQ on the same cell type.
Andreoli
C
C
Istituto Superiore di Sanita', Viale Regina Elena 299, 00161, Rome, Italy.
Rossi
S
S
Leopardi
P
P
Crebelli
R
R
D000328:Adult; D002374:Catalase / Q000494:pharmacology; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005328:Fetal Viability; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D006873:Hydroquinones / Q000633:toxicity*; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*
D006873:Hydroquinones; D009153:Mutagens; D006861:Hydrogen Peroxide; D002374:Catalase; C031927:hydroquinone