Detection of benzo(a)pyrene diol epoxide-DNA adducts in sperm of men exposed to cigarette smoke.
OBJECTIVE
To determine whether the adducts formed when benzo(a)pyrene, a diol epoxide derivative, binds covalently to DNA (BPDE-DNA adducts) are detectable in the sperm of men who smoke cigarettes.


METHODS
Prospective study.


METHODS
The Toronto Hospital IVF-ET program.


METHODS
Twenty-three patients with normal seminal parameters: 11 smokers (20.6 +/- 0.7 cigarettes per day) and 12 nonsmokers.


METHODS
Semen samples obtained by masturbation.


METHODS
Seminal plasma samples were assessed for cotinine by RIA. Sperm were treated with dithiothreitol to release disulfide bonds and allow for DNA binding, then exposed to an anti-BPDE monoclonal antibody, a biotinylated antibody, and streptavidin-conjugated peroxidase. Staining intensity scores, determined in 100 cells per individual, were correlated with seminal plasma cotinine levels, a marker of smoking.


RESULTS
Cotinine levels correlated highly with the number of cigarettes smoked per day. Mean cotinine levels and mean staining intensity scores were higher in smokers than in nonsmokers. Staining intensity correlated highly with cotinine levels.


CONCLUSIONS
We demonstrated, for the first time, that BPDE-DNA adducts in sperm cells are increased by smoking; we also detected comparatively high levels in nonsmokers, which indicates that environmental exposure also is substantial. The formation of adducts in spermatozoa is a potential source of transmissible prezygotic DNA damage.
Zenzes
M T
MT
Toronto Hospital Research Centre, Ontario, Canada.
Bielecki
R
R
Reed
T E
TE
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000032:analysis*; D000328:Adult; D015415:Biomarkers / Q000032:analysis; D003367:Cotinine / Q000032:analysis*; D018736:DNA Adducts / Q000032:analysis*; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D011863:Radioimmunoassay; D012016:Reference Values; D012661:Semen / Q000737:chemistry; D012907:Smoking / Q000503:physiopathology*; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*
D015415:Biomarkers; D018736:DNA Adducts; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D003367:Cotinine