DNA methylation is an epigenetic mechanism regulating transcription, which when disrupted, can alter gene expression and contribute to
carcinogenesis.
Diethanolamine (DEA), a non-genotoxic alkanolamine, produces liver
tumors in mice. Studies suggest DEA inhibits
choline uptake and causes biochemical changes consistent with
choline deficiency (CD). Rodents fed methyl-deficient diets exhibit altered methylation of hepatic
DNA and an increase in liver
tumors, e.g., CD causes liver
tumors in B6C3F1 mice. We hypothesize that DEA-induced CD leads to altered methylation patterns which facilitates
tumorigenesis. B6C3F1 hepatocytes in primary culture were grown in the presence of either 4.5 mM DEA, 3 mM
Phenobarbital (PB), or CD media for 48 h. These concentrations induced comparable increases in
DNA synthesis. PB, a nongenotoxic rodent liver
carcinogen known to alter methylation in mouse liver, was included as a positive control. Global, average, DNA methylation status was not affected. The methylation status of GC-rich regions of
DNA, which are often associated with promoter regions, were assessed via methylation-sensitive restriction digestion and arbitrarily primed PCR with capillary electrophoretic separation and detection of PCR products. DEA, PB, and CD treatments resulted in 54, 63, and 54 regions of altered methylation (RAMs), respectively, and the majority were hypomethylations. A high proportion of RAMs (72%) were identical when DEA was compared to CD. Similarly, 70% were identical between PB and CD. Altered patterns of methylation in GC-rich regions induced by DEA and PB resemble that of CD and indicate that altered DNA methylation is an epigenetic mechanism involved in the facilitation of mouse liver
tumorigenesis.