Synthesis and structure determination of 6-methylbenzo[a]pyrene-deoxyribonucleoside adducts and their identification and quantitation in vitro and in mouse skin.
Activation of the moderate carcinogen 6-methylbenzo[a]pyrene (6-CH(3)BP) by one-electron oxidation to form DNA adducts was studied. Iodine oxidation of 6-CH(3)BP in the presence of dGuo produces BP-6-CH(2)-N(2)dGuo, BP-6-CH(2)-N7Gua and a mixture of 6-CH(3)BP-(1&3)-N7Gua, whereas in the presence of Ade the adducts BP-6-CH(2)-N1Ade, BP-6-CH(2)-N3Ade, BP-6-CH(2)-N7Ade and 6-CH(3)BP-(1&3)-N1Ade are obtained. Furthermore, for the first time an aromatic hydrocarbon radical cation afforded an adduct with dThd, the stable adduct BP-6-CH(2)-N3dThd. Formation of these adducts indicates that the 6-CH(3)BP radical cation has charge localized at the 6, 1 and 3 position. When 6-CH(3)BP was activated by horseradish peroxidase in the presence of DNA, two depurinating adducts were identified, BP-6-CH(2)-N7Gua (48%) and 6-CH(3)BP-(1&3)-N7Gua (23%), with 29% unidentified stable adducts. In the binding of 6-CH(3)BP catalyzed by rat liver microsomes, the same two depurinating adducts, BP-6-CH(2)-N7Gua (22%) and 6-CH(3)BP-(1&3)-N7Gua (10%), were identified, with 68% unidentified stable adducts. In 6-CH(3)BP-treated mouse skin, the two depurinating adducts, BP-6-CH(2)-N7Gua and 6-CH(3)BP-(1&3)-N7Gua, were identified. Although quantitation of these two adducts was not possible due to coelution of metabolites on HPLC, they appeared to be the major adducts found in mouse skin. These results show that 6-CH(3)BP forms depurinating adducts only with the guanine base of DNA, both in vitro and in mouse skin. The weaker reactivity of 6-CH(3)BP radical cation vs. BP radical cation could account for the weaker tumor-initiating activity of 6-CH(3)BP in comparison to that of BP.
Hanson
A A
AA
Eppley Institute for Research in Cancer and Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, NE 68198-6805, USA.
Li
K M
KM
Lin
C H
CH
Jankowiak
R
R
Small
G J
GJ
Rogan
E G
EG
Cavalieri
E L
EL
D000225:Adenine / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001580:Benzopyrenes / Q000737:chemistry* / Q000378:metabolism; D001711:Biotransformation; D002273:Carcinogens / Q000737:chemistry* / Q000378:metabolism; D002417:Cattle; D018736:DNA Adducts / Q000096:biosynthesis / Q000138:chemical synthesis / Q000737:chemistry*; D003854:Deoxyribonucleotides / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D006147:Guanine / Q000737:chemistry / Q000378:metabolism; D007455:Iodine / Q000737:chemistry; D051379:Mice; D010084:Oxidation-Reduction; D012867:Skin / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
D001580:Benzopyrenes; D002273:Carcinogens; D018736:DNA Adducts; D003854:Deoxyribonucleotides; C017503:6-methylbenzo(a)pyrene; D006147:Guanine; D007455:Iodine; D000225:Adenine