Spontaneous and thiazolidinedione-induced B6C3F1 mouse hemangiosarcomas exhibit low ras oncogene mutation frequencies.
Hemangiosarcomasare uncommon malignant endothelial cell tumors in humans and experimental animal species. The mechanisms giving rise to these tumors are poorly understood even though the histotypes are comparable between humans and rodents. Activating mutations in cellular ras protooncogenes have been detected in sporadic and chemically induced human and rodent hemangiosarcomas. Ras activation significantly modulates tumor angiogenesis, suggesting that mutations in ras genes might be causally related to vascular tumorigenesis. To more clearly define the role of ras in experimental vascular tumorigenesis, mutations in the Ki- and Ha-ras genes were characterized in 63 hemangiosarcomas that arose unexpectedly in control and treated B6C3F1 mice during a two-year carcinogenicity study of the thiazolidinedione troglitazone. DNA was extracted from paraffin sections of mouse hemangiosarcomas, control liver, or positive control hepatocellular carcinomas with defined mutations in the Ki- or Ha-ras genes. Exons 1 and 2 of the Ki- and Ha-ras genes were independently amplified using primer extension preamplification/locus-specific heminested PCR, and PCR amplicons were directly sequenced to identify mutations in codons 12, 13, or 61. Activating mutations were detected in 3 of 63 hemangiosarcomas: a single G-->A transition in the second position of Ki-ras codon 13 in a tumor from a treated animal and two G-->T transversions in the second position of Ha-ras codon 13, one in a single tumor from a control animal and one in a tumor from a treated animal. These mutations are consistent with endogenous mutagenesis arising from oxidative DNA damage. The low frequency of mutation (<5%) indicates that ras mutations did not contribute significantly to hemangiosarcoma development and suggests that mutational ras activation may not be a necessary step in vascular tumorigenesis in mice.
Duddy
S K
SK
Parke-Davis Pharmaceutical Research, Warner-Lambert Co., Ann Arbor, Michigan, 48105, USA. Steven.Duddy@wl.com
Gorospe
S M
SM
Bleavins
M R
MR
de la Iglesia
F A
FA
D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D002839:Chromans / Q000633:toxicity*; D004273:DNA, Neoplasm / Q000737:chemistry / Q000302:isolation & purification; D019143:Evolution, Molecular; D005260:Female; D011905:Genes, ras; D006394:Hemangiosarcoma / Q000139:chemically induced* / Q000235:genetics / Q000473:pathology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D017421:Sequence Analysis; D013844:Thiazoles / Q000633:toxicity*; D045162:Thiazolidinediones; D000077288:Troglitazone; D019043:Vascular Neoplasms / Q000139:chemically induced* / Q000235:genetics / Q000473:pathology
D002273:Carcinogens; D002839:Chromans; D004273:DNA, Neoplasm; D013844:Thiazoles; D045162:Thiazolidinediones; D000077288:Troglitazone