Mechanistic linkage between DNA adducts, mutations in oncogenes and tumorigenesis of carcinogenic environmental polycyclic aromatic hydrocarbons in strain A/J mice.
Five polycyclic aromatic hydrocarbons (PAHs), benzo[a]pyrene (B[a]P), benzo[b]fluoranthene (B[b]F), dibenz[a,h]anthracene (DBA), 5-methylchyrsene (5MC), and cyclopenta[cd]pyrene (CPP) were examined for their lung tumorigenic activities in strain A/J mice, their ability to form PAH-DNA adducts in lung tissues, and their ability to mutate the Ki-ras oncogene in PAH-induced tumors. PAHs dissolved in tricapyrlin were administered by single intraperitoneal injection to male strain A/J mice (20 mice/dose) at doses up to 200 mg/kg depending on the PAH. Animals were sacrificed 8 months later and the lungs removed, fixed, and surface adenomas enumerated. DBA produced maximal tumor multiplicity at the highest dose, 10 mg/kg, giving 32.2 lung adenomas per mouse. At 100 mg/kg, B[a]P, B[b]F, 5MC, and CPP gave 12.8, 5.3, 93.1, and 32.2 lung adenomas per mouse, respectively. The dose response data for each PAH was fit to y = 0.6 + bx1.6, where y is the observed mean lung adenomas per mouse at dose x (in mg/kg), 0.6 is the observed background of lung adenomas per mouse, and b is the fitted constant representing the potency of each PAH. Statistical analysis indicated that the fit of the data to the equation was extremely high with adjusted R2 values > 0.985 and small fit standard errors. Based on this equation, the relative potencies of B[b]F, DBA, 5MC, and CPP compared to B[a]P were PAH (relative activity): DBA (118); 5MC (8.8); CPP (2.9); B[a]P (1.0); B[b]F (0.43). DNA adducts were measured by 32P-postlabeling techniques on DNA from lungs of mice treated with these PAHs. Adducts identified by cochromatography with standards were: from B[a]P, 7R,8S,9S-trihydroxy-10R-(N2-2'-deoxyguanosyl)-7,8,9,10-tetrahydro- B[a]P, and two adducts resulting from the metabolic activation of 9-hydroxy-B[a]P and trans-7,8-dihydroxy-7,8-dihydro-B[a]P; from B[b]F, 5-hydroxy-B[b]F-9,10-diol-11,12-oxide-2'-deoxyguanosine; from DBA, three adducts from the metabolic activation of trans,trans-3,4,10,11-tetrahydroxy-3,4,10,11-tetrahydro-DBA and two anti-DBA-3,4-diol-1,2-oxide-N2-[2'-deoxyguanosine] adducts; from 5MC, 1R,2S,3S-trihydroxy-4-(N2-2'-deoxyguanosyl)-1,2,3,4-tetrahydro- 5MC; from CPP, four CPP-3,4-oxide-2'-deoxyguanosine adducts. Ki-ras codon 12 mutation analysis of PAH-induced tumors was performed using PCR and dideoxy sequencing methods. Mutations from lung tumors from tricaprylin-treated mice were GGT-->GAT, GGT-->CGT, and GGT-->GTT. DBA produced no mutations in Ki-ras codon 12 above spontaneous levels. High proportions (> or = 50%) of GGT-->TGT mutations from B[a]P, B[b]F and 5MC induced tumors and GGT-->CGT mutations from CPP tumors were observed and were statistically significant compared to mutations in tricaprylin control tumors. We conclude from the DNA adduct and Ki-ras mutation studies that bay region diol-epoxide-2'-deoxyguanosine PAH-DNA adducts are associated with the GGT-->TGT mutations, and cyclopenta-ring oxide-2'-deoxyguanosine adducts associated with the GGT-->CGT mutations.
Nesnow
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Carcinogenesis and Metabolism Branch (MD-68), US Environmental Protection Agency, Research Triangle Park, NC 27711, USA.
Ross
J A
JA
Stoner
G D
GD
Mass
M J
MJ
D000236:Adenoma / Q000139:chemically induced / Q000235:genetics / Q000473:pathology; D000818:Animals; D001483:Base Sequence; D001711:Biotransformation; D002273:Carcinogens / Q000633:toxicity*; D018736:DNA Adducts / Q000096:biosynthesis*; D011905:Genes, ras / Q000235:genetics*; D008175:Lung Neoplasms / Q000139:chemically induced / Q000235:genetics / Q000473:pathology; D008297:Male; D051379:Mice; D008805:Mice, Inbred A; D008969:Molecular Sequence Data; D009154:Mutation / Q000187:drug effects; D009369:Neoplasms / Q000139:chemically induced*; D011084:Polycyclic Aromatic Hydrocarbons / Q000737:chemistry / Q000493:pharmacokinetics / Q000633:toxicity*; D016133:Polymerase Chain Reaction; D013329:Structure-Activity Relationship
D002273:Carcinogens; D018736:DNA Adducts; D011084:Polycyclic Aromatic Hydrocarbons