A short-term assessment of tumor-promotion activity in the livers of rats treated with two genotoxic methylating agents: dimethylnitrosamine and methylnitrosourea.
Induction of replicative DNA synthesis (RDS) and mitoinhibitory effects were studied in the hepatocytes of F344 rats exposed in vivo to the methylating agents dimethylnitrosamine (DMN, hepatocarcinogen) and methylnitrosourea (MNU, non-hepatocarcinogen). Cytotoxicity and chromosome aberrations (CA) in rat liver were also investigated to clarify the cause of changes in RDS and mitoinhibitory effects, respectively. The animals were killed at different intervals (up to 14 days) after a single oral dose, or 1 day after 7 or 14 days of repeated oral doses. The hepatocytes were isolated and cultured with Williams' medium E to assess their RDS, mitoinhibitory effects and CA. Mitoinhibitory effects were investigated by monitoring their effect on epidermal growth factor-induced replicative DNA synthesis (EGF-induced RDS) in rat hepatocytes. Hepatotoxic effects were assessed by measuring aspartate transaminase and alanine transaminase in the plasma and by histopathological examination. In the single-dose study, DMN (20 mg/kg body weight (bw)) induced both RDS and hepatotoxicity. MNU (50 mg/kg bw) induced RDS without causing hepatotoxicity, and thus was classified as a mitogen. In the repeated-dose study, DMN (4 mg/kg bw) induced both RDS and hepatotoxicity, but MNU (10 mg/kg bw) induced neither. Both inhibition of EGF-induced RDS and induction of CA were observed in the hepatocytes of rats treated with DMN, but were not observed with MNU in both single and repeated dose studies. The mitoinhibitory effect of DMN persisted for 14 days after the single dose and time dependently increased for 14 days after repeated administration. This mitoinhibitory effect correlated positively with CA. The mitoinhibitory effect was thought to be attributable to the DNA-damaging effect that induces CA. We concluded that the differences which we found in this study between DMN and MNU contribute to the differences in their hepatocarcinogenicity. Our findings suggested that both cell proliferative and mitoinhibitory properties play an important role in tumor promotion, and measuring them may provide an ancillary index that is useful in predicting hepatocarcinogenicity.
Asakura
S
S
Tsukuba Drug Safety Research, Drug Safety Research Laboratories, Eisai Co., Ltd., Ibaraki, Japan. s1-asakura@eisai.co.jp
Daimon
H
H
Sawada
S
S
Sagami
F
F
D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D002273:Carcinogens / Q000633:toxicity*; D002455:Cell Division / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D004261:DNA Replication / Q000187:drug effects; D004128:Dimethylnitrosamine / Q000633:toxicity*; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D008114:Liver Neoplasms, Experimental / Q000139:chemically induced* / Q000201:enzymology / Q000473:pathology; D008297:Male; D008770:Methylnitrosourea / Q000633:toxicity*; D051381:Rats; D011916:Rats, Inbred F344
D002273:Carcinogens; D008770:Methylnitrosourea; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D004128:Dimethylnitrosamine