Mutagenesis of ras proto-oncogenes in rat liver tumors induced by vinyl chloride.
Vinyl chloride is a DNA-damaging carcinogen which induces liver angiosarcomas in humans and animals. Activation of the Ki-ras 2 gene by a GC-->AT transition at the second base of codon 13 in human liver angiosarcomas associated with occupational exposure to vinyl chloride has been reported recently. In order to compare the molecular pathways of carcinogenesis in humans and animals, Sprague-Dawley rats were exposed to vinyl chloride and hepatic tumors, including two hepatocellular carcinomas and five liver angiosarcomas, were investigated for mutations at codons 12, 13 and 61 of the Ha-ras, Ki-ras and N-ras genes. High molecular weight DNA was amplified by the polymerase chain reaction and point mutations were analyzed by allele specific oligonucleotide hybridization, direct sequencing of polymerase chain reaction products and sequencing after cloning. None of the tumors exhibited a mutation in codons 12, 13 and 61 of the Ki-ras gene, nor in codons 12 of the Ha-ras gene or 61 of the N-ras gene. However, an activating AT-->TA transversion at base 2 of codon 61 of the Ha-ras gene was detected in the two hepatocellular carcinomas. Mutations involving codon 13 (GGC-->GAC) and codon 36 (ATA-->CTA) of the N-ras A gene were detected in two liver angiosarcomas, suggesting that the nature of the ras gene affected by a given carcinogen depends on host factors specific to cell types. Several additional base pair substitutions were found in exon 1 of the N-ras B and C sequences. NIH 3T3 transfection assays and Southern blot analysis of DNA from transformed NIH 3T3 cells confirmed the presence of a dominant activated N-ras gene. These results emphasize the differences in the molecular pathways leading to tumors in humans and rats and within a given species between different cell types.
Froment
O
O
Institut National de la Santé et de la Recherche Médicale, Unité 271, Hepatitis Research Unit, Lyon, France.
Boivin
S
S
Barbin
A
A
Bancel
B
B
Trepo
C
C
Marion
M J
MJ
D016475:3T3 Cells; D002759:Adenoma, Bile Duct / Q000139:chemically induced / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D003062:Codon / Q000235:genetics*; D005260:Female; D011905:Genes, ras / Q000235:genetics*; D006394:Hemangiosarcoma / Q000139:chemically induced / Q000235:genetics*; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced / Q000235:genetics*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D017354:Point Mutation; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D014752:Vinyl Chloride
D003062:Codon; D014752:Vinyl Chloride