DNA adducts, strand breaks and micronuclei in mice exposed to styrene by inhalation.
Genotoxic and clastogenic effects of styrene were studied in mice. Male NMRI mice were exposed by inhalation to styrene in concentrations of 750 and 1500 mg/m3 for 21, 7, 3 and 1 days (6 h/day, 7 days/week). Followed parameters included styrene in blood, specific styrene oxide (SO) induced DNA adducts, DNA strand breaks and micronuclei. The formation of SO induced 7-SO-guanines and 1-SO-adenines in DNA was analysed from lung tissues by two versions of the 32P-postlabeling technique. In lungs after 21 days of exposure to 1500 mg/m3 the level of 7-SO-guanine was 23.0+/-11.9 adducts/10(8) normal nucleotides, while 1-SO-adenine was detected at the levels of 0.6+/-0.2 adducts/10(8) normal nucleotides. Both 7-SO-guanines and 1-SO-adenines strongly correlated with exposure parameters, particularly with styrene concentration in blood (r=0.875, P=0.0002 and r=0.793, P=0.002, respectively). DNA breaks were measured in peripheral lymphocytes, bone marrow cells and liver cells using comet assay. To discern oxidative damage and abasic sites, endonuclease III was used. In bone marrow of exposed mice slight increase of strand breaks can be detected after 7 days of inhalation. A significant increase was revealed in the endonuclease III-sensitive sites after 21 days of inhalation in bone marrow. In the liver cells inhalation exposure to both concentrations of styrene did not virtually affect either levels of DNA single-strand breaks or endonuclease III-sensitive sites. The inhalation of 1500 mg/m3 of styrene induced significant increase of micronuclei after 7 days of exposure (10.4+/-2.5/1000 cells, i.e. twice higher micronuclei frequency than in controls). After 21 days of inhalation no significant difference between the control group and the two exposed groups was observed. Whether the decrease of micronuclei after 21 days of inhalation was due to the inhibition of cell proliferation caused by styrene or due to the natural elimination of chromatide fragments, remains to be clarified. An interesting link has been found between DNA single-strand breaks in bone marrow and frequencies of micronuclei (r=0.721, P=0.028).
Vodicka
P
P
Institute of Experimental Medicine, Academy of Sciences of Czech Republic, Videnska 1083, 14220 4, Prague, Czech Republic. pvodicka@biomed.cas.cz
Koskinen
M
M
Vodicková
L
L
Stetina
R
R
Smerák
P
P
Bárta
I
I
Hemminki
K
K
D000280:Administration, Inhalation; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D020552:Comet Assay; D004247:DNA / Q000187:drug effects*; D018736:DNA Adducts / Q000378:metabolism*; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004852:Epoxy Compounds / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008214:Lymphocytes / Q000187:drug effects; D008297:Male; D051379:Mice; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects; D009153:Mutagens / Q000633:toxicity*; D020058:Styrene / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000633:toxicity*
D018736:DNA Adducts; D004852:Epoxy Compounds; D009153:Mutagens; D020058:Styrene; D004247:DNA; C013690:styrene oxide