Etheno-adduct-forming chemicals: from mutagenicity testing to tumor mutation spectra

During the past 25 years, ethenobases have emerged as a new class of DNA lesions with promutagenic potential. Ethenobases were first investigated as DNA reaction products of vinyl chloride, an occupational carcinogen causing angiosarcoma of the liver (ASL). They were subsequently shown to be formed by several carcinogenic agents, including urethane (ethyl carbamate), and more recently, to occur in various tissues of unexposed humans and rodents. The endogenous source of ethenobases in DNA is thought to be a lipid peroxidation (LPO) product. Initial studies on metabolic activation, mutagenicity and carcinogenicity moved to the analyses of the formation of ethenobases in vivo and to the determination of their promutagenic properties. Quantification of etheno adducts in vivo became possible with the development of ultrasensitive techniques of analysis. To study the miscoding properties of ethenobases, the initial assays on the fidelity of replication or of transcription were replaced by site-directed mutagenesis assays in vivo. Ethenobases generate mainly base pair substitution mutations. With the advent of new techniques of molecular biology, mutations were investigated in the ras and p53 genes of tumors induced by vinyl chloride and urethane. In liver tumors induced by vinyl chloride, specific mutational patterns were found in the Ki-ras gene in human ASL, in the Ha-ras gene in hepatocellular carcinoma (HCC) in rats, and in the p53 gene in human and rat ASL. In tumors induced by urethane in mice, codon 61 of the Ha-ras gene (liver, skin) and of the Ki-ras gene (lung) seems to be a characteristic target. These tumor mutation spectra are compatible with the promutagenic properties of etheno adducts and with their formation in target tissues, suggesting that ethenobases can be initiating lesions in carcinogenesis. Another recent focus has been given to the repair of etheno adducts, and DNA glycosylases able to excise these adducts in vitro have been identified. The last two decades have brought ethenobases to light as potentially important DNA lesions in carcinogenesis. More research is needed to better understand the environmental and genetic factors that affect the formation and persistence of ethenobases in vivo.