Wy-14,643-induced hypomethylation of the c-myc gene in mouse liver.

The carcinogenic activity of Wy-14,643 in mouse liver appears to be nongenotoxic and could involve a decrease in DNA methylation. The mechanism for Wy-14,643-induced decrease in DNA methylation is proposed to involve increased cell proliferation followed by prevention of the methylation of the newly synthesized DNA. To investigate this mechanism, female B6C3F1 mice were administered daily by oral gavage 50 mg/kg Wy-14,643. Mice were sacrificed at 2, 5, 8, 24, 26, 29, 32, 36, 48, 72, and 96 h after the first dose. Some mice also received 450 mg/kg methionine by ip injection at 30 min after administering Wy-14,643. Hypomethylation of the c-myc gene first occurred at 48 h after the first dose of Wy-14,643. Cell proliferation determined by the Proliferating Cell Nuclear Antigen (PCNA)-Labeling Index started to increase at 36 h and peaked at 72h. Wy14,643 did not affect the liver concentration of either S-adenosyl methionine (SAM) or S-adenosyl homocysteine (SAH). Methionine prevented and reversed the hypomethylation of the c-myc gene induced by Wy-14,643. However, the increased levels of SAM and SAH returned to control levels prior to the prevention by methionine of Wy-14,643-induced hypomethylation. Furthermore, methionine did not prevent Wy-14,643-induced increase in the PCNA-Labeling Index. The activity of nuclear DNA methyltransferase (DNA MTase) was increased at 72 and 96 h after administering Wy14,643. Wy14,643 also increased the activity of DNA MTase when added in vitro to nuclear extracts. The results are consistent with Wy-14,643 decreasing the methylation of the c-myc gene by a mechanism that includes enhancement of cell proliferation followed by prevention of the methylation of the newly synthesized DNA. However, the results indicate that Wy-14,643 does not prevent methylation by decreasing either the availability of SAM or the activity of DNA MTase.