Structures and biological properties of DNA adducts derived from N-nitroso bile acid conjugates.
A kind of N-nitrosobile acid conjugate, N-nitrosotaurocholic acid (NO-TCA), was incubated with calf thymus DNA, and formation of an adduct was detected by the 32P-postlabeling method under nuclease P1 conditions. To examine the nucleotides containing the adduct from NO-TCA, each of 2'-deoxyribonucleotide 3'-monophosphates (3'-dAp, 3'-dGp, 3'-dCp, or 3'-Tp) was incubated with NO-TCA. The same adduct spot was detected in the reaction of NO-TCA with 3'-dCp. The structure of this adduct was determined to be 3-ethanesulfonic acid-dC by several spectrometry techniques. Moreover, bulky adducts containing bile acid moiety were also produced from the reaction of NO-TCA with 3'-dCp and 3'-dAp. From comparison with spectral data for authentic compounds, these adducts were concluded to be N4-cholyl-dC and N6-cholyl-dA. N4-Cholyl-dC and N6-cholyl-dA were also detected in calf thymus DNA treated with NO-TCA. In addition, 3-ethanesulfonic acid-dC and N4-deoxycholyl-dC were found to be produced from N-nitrosotaurodeoxycholic acid (NO-TDCA) with dC. NO-TCA and NO-TDCA induced mutations in Salmonella typhimurium TA100 but not in TA98. Mutational spectrum analysis revealed that NO-TCA induced G to A transitions predominantly. When NO-TCA (250 mg/kg) was singly administered to male Wistar rats by gavage, both ethanesulfonic acid-dC and N4-cholyl-dC could be detected in the glandular stomach and colon. The levels of ethanesulfonic acid-dC were 0.22-0.29 per 10(6) nucleotides, but values for N4-cholyl-dC were about 500-fold lower. These observations suggest that N-nitroso bile acid conjugates, NO-TCA and NO-TDCA, may induce G to A base substitutions in genes via DNA adduct formation, producing ethanesulfonic acid- and/or (deoxy)cholic acid-DNA and, therefore, may be related to human carcinogenesis as endogenous mutagens.
Totsuka
Yukari
Y
Cancer Prevention Basic Research Project, National Cancer Center Research Institute, 1-1 Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan. ytotsuka@gan2.res.ncc.go.jp
Nishigaki
Rena
R
Enomoto
Shigeki
S
Takamura-Enya
Takeji
T
Masumura
Ken-ichi
K
Nohmi
Takehiko
T
Kawahara
Nobuo
N
Sugimura
Takashi
T
Wakabayashi
Keiji
K
D000476:Alkanesulfonates / Q000737:chemistry; D000818:Animals; D001647:Bile Acids and Salts / Q000737:chemistry* / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry* / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000378:metabolism; D008297:Male; D009153:Mutagens / Q000633:toxicity; D009154:Mutation; D009602:Nitrosamines / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D013657:Taurodeoxycholic Acid / Q000737:chemistry / Q000378:metabolism
D000476:Alkanesulfonates; D001647:Bile Acids and Salts; D018736:DNA Adducts; D009153:Mutagens; D009602:Nitrosamines; D013657:Taurodeoxycholic Acid; C002634:ethane sulfonate; C034936:N-nitrosotaurocholic acid