Processing of DNA damage induced by hydrogen peroxide and methyl methanesulfonate in human lymphocytes: analysis by alkaline single cell gel electrophoresis and cytogenetic methods

被引:32
作者
Andreoli, C [1 ]
Leopardi, P [1 ]
Rossi, S [1 ]
Crebelli, R [1 ]
机构
[1] Ist Super Sanita, I-00161 Rome, Italy
关键词
D O I
10.1093/mutage/14.5.497
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The persistence of induced DNA damage in human lymphocytes after mitogen stimulation and its relationship to subsequent cytogenetic alterations were investigated. The analysis of single-strand breaks and alkali-labile sites by single cell gel electrophoresis (SCGE) showed the almost complete repair of damage induced in resting lymphocytes by methyl methanesulfonate (MMS, 140-210 mu M) and hydrogen peroxide (H2O2, 25-100 mu M) during the first 16 h of culture. On the other hand, DNA damage was shown to persist to a large extent when cells were cultured in the presence of the repair inhibitor cytosine beta-D-arabinofuranoside (Ara-C) (1 mu g/ml). Although highly effective in the induction of DNA lesions detectable by SCGE, both agents failed to significantly increase the rate of micronucleus formation in cytokinesis-blocked cells harvested 66 h after treatment. However, when Ara-C was present during the first 16 h of culture, micronuclei were significantly increased at all doses. Conversely, sister chromatid exchange (SCE) rates were increased by chemical treatments to a higher extent in cultures without Ara-C, Delayed treatments, 16 h after mitogen stimulation, led to a significant induction of micronuclei in the case of MMS but not with H2O2. These results suggest that only a minor fraction of DNA damage induced in resting lymphocytes is available for fixation through misreplication, because of its effective repair prior to S phase. However, the processing of damage through recombination pathways can lead to increased SCE rates in treated cells, These features of the processing of DNA damage in human lymphocytes should be taken into account when structural cytogenetic alterations in cultured lymphocytes are used in monitoring human exposure to genotoxic agents.
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页码:497 / 503
页数:7
相关论文
共 42 条
[1]   Detection of DNA damage in human lymphocytes by alkaline single cell gel electrophoresis after exposure to benzene or benzene metabolites [J].
Andreoli, C ;
Leopardi, P ;
Crebelli, R .
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 1997, 377 (01) :95-104
[2]   DNA damage by hydroquinone in human white blood cells: analysis by alkaline single-cell gel electrophoresis [J].
Andreoli, C ;
Rossi, S ;
Leopardi, P ;
Crebelli, R .
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS, 1999, 438 (01) :37-45
[3]   DISTRIBUTION OF METHYL AND ETHYL ADDUCTS FOLLOWING ALKYLATION WITH MONOFUNCTIONAL ALKYLATING-AGENTS [J].
BERANEK, DT .
MUTATION RESEARCH, 1990, 231 (01) :11-30
[4]   ASSESSMENT OF DNA-DAMAGE IN LEUKOCYTES FROM INFECTED AND MALNOURISHED CHILDREN BY SINGLE-CELL GEL-ELECTROPHORESIS COMET ASSAY [J].
BETANCOURT, M ;
ORTIZ, R ;
GONZALEZ, C ;
PEREZ, P ;
CORTES, L ;
RODRIGUEZ, L ;
VILLASENOR, L .
MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 1995, 331 (01) :65-77
[5]   COMPARATIVE-STUDIES BY COMET TEST AND SCE ANALYSIS IN HUMAN-LYMPHOCYTES FROM 200 HEALTHY-SUBJECTS [J].
BETTI, C ;
DAVINI, T ;
GIANNESSI, L ;
LOPRIENO, N ;
BARALE, R .
MUTATION RESEARCH-GENETIC TOXICOLOGY, 1995, 343 (04) :201-207
[6]   Biomarker studies in northern Bohemia [J].
Binkova, B ;
Lewtas, J ;
Miskova, I ;
Rossner, P ;
Cerna, M ;
Mrackova, G ;
Peterkova, K ;
Mumford, J ;
Meyer, S ;
Sram, R .
ENVIRONMENTAL HEALTH PERSPECTIVES, 1996, 104 :591-597
[7]  
Collins A, 1997, ENVIRON MOL MUTAGEN, V30, P139, DOI 10.1002/(SICI)1098-2280(1997)30:2<139::AID-EM6>3.0.CO
[8]  
2-I
[9]   DIRECT ENZYMATIC DETECTION OF ENDOGENOUS OXIDATIVE BASE DAMAGE IN HUMAN LYMPHOCYTE DNA [J].
COLLINS, AR ;
DUTHIE, SJ ;
DOBSON, VL .
CARCINOGENESIS, 1993, 14 (09) :1733-1735
[10]   THE KINETICS OF REPAIR OF OXIDATIVE DNA-DAMAGE (STRAND BREAKS AND OXIDIZED PYRIMIDINES) IN HUMAN-CELLS [J].
COLLINS, AR ;
MA, AG ;
DUTHIE, SJ .
MUTATION RESEARCH-DNA REPAIR, 1995, 336 (01) :69-77