Mutagenicity of carcinogenic heterocyclic amines in Salmonella typhimurium YG strains and transgenic rodents including gpt delta

Chemical carcinogens to humans have been usually identified by epidemiological studies on the relationships between occupational or environmental exposure to the agents and specific cancer induction. In contrast, carcinogenic heterocyclic amines were identified under the principle that mutagens in bacterial in the Ames test are possible human carcinogens. In the 1970s to 1990s, more than 10 heterocyclic amines were isolated from pyrolysates of amino acids, proteins, meat or fish as mutagens in the Ames test, and they were demonstrated as carcinogens in rodents. In the 1980s and 1990s, we have developed derivatives of the Ames tester strains that overexpressed acetyltransferase of Salmonella typhimurium. These strains such as Salmonella typhimurium YG1024 exhibited a high sensitivity to the mutagenicity of the carcinogenic heterocyclic amines. Because of the high sensitivity, YG1024 and other YG strains were used for various purposes, e.g., identification of novel heterocyclic amines, mechanisms of metabolic activation, comparison of mutagenic potencies of various heterocyclic amines, and the co-mutagenic effects. In the 1990s and 2000s, we developed transgenic mice and rats for the detection of mutagenicity of chemicals in vivo. The transgenics were generated by the introduction of reporter genes for mutations into fertilized eggs of mice and rats. We named the transgenics as gpt delta because the gpt gene of Escherichia coli was used for detection of point mutations such as base substitutions and frameshifts and the red/gam genes of λ phage were employed to detect deletion mutations. The transgenic rodents gpt delta and other transgenics with lacI or lacZ as reporter genes have been utilized for characterization of mutagenicity of heterocyclic amines in vivo. In this review, we summarized the in vitro mutagenicity of heterocyclic amines in Salmonella typhimurium YG strains and the in vivo mutagenicity in transgenic rodents. We discussed the relationships between in vitro and in vivo mutagenicity of the heterocyclic amines and their relations to the carcinogenicity.

Lack of In Vivo Mutagenicity of Acetamide in a 13-Week Comprehensive Toxicity Study Using F344 gpt Delta Rats

Acetamide, a food contaminant, has been shown to induce hepatocellular tumors in rats. However, the mode of action underlying acetamide-induced hepatocarcinogenesis remains unclear. In the current study, we aimed to examine the possible involvement of in vivo mutagenicity in hepatocarcinogenesis of acetamide and evaluate its toxicological profile using a comprehensive medium-term toxicity study in gpt delta rats. Six-week-old male F344 gpt delta rats were given a basal diet containing 0%, 0.625%, 1.25%, or 2.5% acetamide for 13 weeks. In general toxicologic assessment, hepatotoxic parameters in serum, such as aspartate aminotransferase and alanine aminotransferase were significantly changed at the 1.25% group and higher. Histopathological examination of the liver revealed that various changes related to hepatic injury were observed at the 1.25% group and higher. Interestingly, Feulgen-positive cytoplasmic inclusion was frequently observed in hepatocytes in these groups. In the hematopoietic system, red blood cell parameters in plasma, such as mean corpuscular volume and mean corpuscular hemoglobin were significantly changed at the 1.25% group and higher, and decrease of erythroblast in the spleen was observed histopathologically in the 2.5% group. Thus, the no-observed-adverse-effect level of acetamide in this study was 0.625% (equivalent to 394 mg/kg body weight/day). In vivo mutation assays showed that acetamide induced no changes in gpt and red/gam gene mutant frequencies, even at the carcinogenic target site. In contrast, Ki67-positive hepatocytes were increased significantly at carcinogenic doses. Therefore, these results suggested that cell proliferation activity, but not mutagenicity, played crucial roles in acetamide-induced hepatocarcinogenesis in rats.