title: H-ras 61st codon activation in archival proliferative hepatic lesions isolated from female B6C3F1 mice exposed to the leukotriene D4-antagonist, LY171883.
abstract: LY171883, a peroxisome proliferator and leukotriene D4-antagonist, induced a statistically significant increase in the number of hepatic lesions in B6C3F1 female mice in a 2 year oncogenicity study at dietary doses of 0.0225% and 0.075%. The mutation frequency and spectrum of the 61st codon of H-ras was determined for 64 independent, archived lesions from the LY171883 2 year oncogenicity study using the polymerase chain reaction (PCR), allele specific oligo hybridization (ASO) and DNA sequencing. Results showed 41 (64%) of these lesions had mutations at the 61st codon (16/21 hepatocellular carcinomas, 4/10 hepatocellular adenomas, 19/26 focal hepatocellular hyperplasias and 2/7 focal hepatocellular atypia). These mutations consisted of 18 C-A transversions, 16 A-G transitions and seven A-T transversions. Compared to the mutation frequency for spontaneously occurring archival B6C3F1 hepatic lesions (41%), the frequency of LY171883 lesions (64%) was significantly higher (P < 0.01). The frequencies of H-ras 61st codon mutations among the LY171883 lesion types (hepatocellular carcinomas 76%, hepatocellular adenomas 40%, focal hepatocellular hyperplasias 73% and hepatocellular atypia 29%) were also significantly different (P = 0.035). In contrast, spontaneous lesions showed no statistical difference in the frequencies of mutation among lesion types (P > 0.5). The mutation spectrum of the LY171883 lesions was not significantly different from the spontaneous spectra. It may be concluded that based on the similarity in mutation spectrum and the increase in mutation frequency, LY171883 may selectively promote spontaneous hepatic lesions containing H-ras 61st codon mutations. In addition, the difference in mutation frequency among lesion types does not support a linear progression of all LY171883 lesions through focal atypia, focal hepatocellular hyperplasias, hepatocellular adenomas and hepatocellular carcinomas.
authors.affiliation: Toxicology Research Laboratories, Lilly Research Laboratories, Greenfield, IN 46140.
authors.forename: L M
authors.initials: LM
authors.lastname: Helvering
authors.forename: F C
authors.initials: FC
authors.lastname: Richardson
authors.forename: D M
authors.initials: DM
authors.lastname: Horn
authors.forename: M A
authors.initials: MA
authors.lastname: Rexroat
authors.forename: J A
authors.initials: JA
authors.lastname: Engelhardt
authors.forename: K K
authors.initials: KK
authors.lastname: Richardson
mesh_terms: D000098:Acetophenones / Q000633:toxicity*; D000818:Animals; D001483:Base Sequence; D002273:Carcinogens / Q000633:toxicity*; D003062:Codon; D017931:DNA Primers; D005260:Female; D011905:Genes, ras; D017998:Leukotriene D4 / Q000037:antagonists & inhibitors*; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D013777:Tetrazoles / Q000633:toxicity*
chemical_list: D000098:Acetophenones; D002273:Carcinogens; D003062:Codon; D017931:DNA Primers; D013777:Tetrazoles; D017998:Leukotriene D4; C044660:LY 171883