5-methyldeoxycytidine and Liver-Neoplasms

Cellular methylation imbalance is associated with tumor progression, hepatic cancer, and cardiovascular disease. S-Adenosylhomocysteine (SAH) is an inhibitor of cellular methyltransferases, and increasing evidence suggests that SAH rather than homocysteine (Hcy) plays a crucial role in mediating these disorders related to methylation imbalance. The anti-metastatic gene nm23-H1 was recently identified in murine and human cancer lines, and the expressions of nm23-H1 mRNA and protein have been shown to be useful tumor invasion markers. We investigated the relationships of tumor cell invasion activities with the intracellular levels of SAH and Hcy and the level of DNA methylation (measured as the cellular content of 5-methyldeoxycytidine, 5-mdc) in four hepatocarcinoma cell lines (Sk-Hep1, J5, Hep-G2, Hep-3B) and one normal liver cell line (Chang's liver cells) with different invasion activities (Sk-Hep1 > J5 > Hep-G2 = Hep-3B > Chang's liver cells). We found that the intracellular level of SAH was the highest in SK-Hep1 cells and was correlated with the invasion activities (r = 0.75, P = 0.008), whereas the level of intracellular Hcy was the highest in Chang's liver cells and was not significantly correlated with the invasion activities of these cell lines (r = 0.24, P = 0.38). The levels of 5-mdc increased with decreasing invasion activities of these cell lines (r = 0.82, P = 0.002), that is, the order of DNA hypomethylation in these cell lines was Sk-Hep1 > J5 > Hep-G2 = Hep-3B > Chang's liver cells, because the lower levels of 5-mdc% represent the higher DNA hypomethylation. Thus, our results demonstrate that SAH rather than Hcy is associated with invasion activities of hepatoma cells, and they suggest that SAH may play an important role in the invasion activities through DNA hypomethylation.

Topics: Cell Line, Tumor; Deoxycytidine; DNA Methylation; Down-Regulation; Homocysteine; Humans; Liver Neoplasms; Neoplasm Invasiveness; S-Adenosylhomocysteine

The protooncogene c-myc was investigated in N-nitrosomorpholine-induced rat liver nodules to elucidate the role of altered DNA methylation in chemical carcinogenesis. Furthermore, Micrococcus luteus DNA and chicken erythrocyte DNA were modified in vitro by reactive metabolites of N-nitrosomorpholine, generated by P450-dependent monooxygenases. The modified DNAs were less methylated in vitro than control DNAs by DNA-(cytosine-5)-methyltransferase (DNA methylase). The DNA methylase assay and 32P-postlabeling analysis revealed lowered levels of DNA methylation in nodular DNA. In nodular tissue, c-myc messenger RNA levels were found to be increased compared to normal liver. DNA methylation analysis using the restriction endonucleases HpaII/MspI indicated hypomethylation in the first intron of c-myc DNA in liver nodules. The results suggest that genotoxic lesions may cause stably inherited, aberrant DNA methylation patterns which may be responsible for site-specific hypomethylation of the c-myc protooncogene in liver nodules.

Topics: Animals; Deoxycytidine; DNA Modification Methylases; DNA, Bacterial; DNA, Neoplasm; Liver Neoplasms; Male; Methylation; Nitrosamines; Precancerous Conditions; Proto-Oncogenes; Rats; Rats, Inbred Strains; RNA, Neoplasm

The effects of the chronic administration of methyl-deficient, amino acid-defined diets on liver tumor formation were examined in male weanling C3H/HeN mice previously treated with a single ip injection of 0 or 150 mg diethylnitrosamine/kg body weight [(DENA) CAS: 55-18-5]. Five diets were used: diet 1, adequate; diet 2, devoid of both methionine and choline; diet 3, devoid of methionine only; diet 4, devoid of choline only; and diet 5, devoid of methionine, choline, folic acid, and vitamin B12. Equimolar homocystine replaced methionine in all methionine-devoid diets. All diets were administered for 1 year. No hepatocellular carcinomas and only 3 liver adenomas were seen among the 129 animals at risk in the 5 groups that had received no DENA. Among the DENA-treated groups fed diets 1-4, the incidence of hepatocellular carcinomas in the mice at risk averaged 40%, with no significant differences noted among groups. A relatively low incidence of liver carcinomas (10%) was seen among DENA-treated mice subsequently fed diet 5; it could be ascribed to the enhanced mortality seen in these animals due to the dietary deficiencies. Lung tumors were seen in 44% of the DENA-treated mice surviving more than 35 experimental weeks and in only 2.5% of the corresponding DENA-untreated animals. Feeding diet 2, deficient in methionine and choline, to male C3H mice for 5-20 weeks decreased the hepatic ratio of S-adenosylmethionine (CAS: 29908-3-0) to S-adenosylhomocysteine (979-92-0) relative to that observed in mice fed the adequate diet 1. The 5-methyldeoxycytidine [(5-MC) CAS: 838-07-3] contents of liver DNA in animals fed diet 2 for 5, 10, and 20 weeks, however, were not significantly different from the corresponding levels in diet 1-fed mice. The results indicate that a methionine- and choline-deficient dietary regimen that lowers the 5-MC levels in DNA and enhances liver tumor formation in male F344 rats does not do so in male C3H mice.

Topics: Animals; Body Weight; Choline; Deoxycytidine; Diet; Diethylnitrosamine; DNA; Liver; Liver Neoplasms; Male; Methionine; Methylation; Mice; Mice, Inbred C3H; Organ Size; S-Adenosylhomocysteine; S-Adenosylmethionine; Time Factors

A progressive decrease was observed in the 5-methyldeoxycytidine content of hepatic DNA in male F344 rats fed with a hepatocarcinogenic methyl-deficient diet. The same dietary regimen resulted in altered hepatic contents of S-adenosylmethionine, the methyl-donating species, and S-adenosylhomocysteine, an inhibitor of DNA methylase. The data indicate that this carcinogenic dietary manipulation is sufficient to alter a possible regulatory process, DNA methylation.

Topics: Animals; Cell Nucleus; Choline Deficiency; Deoxycytidine; Diet; Diethylnitrosamine; DNA; Liver; Liver Neoplasms; Male; Methionine; Methylation; Rats; Rats, Inbred F344; S-Adenosylhomocysteine; S-Adenosylmethionine