The value of DNA methylation analysis in basic, initial toxicity assessments.
DNA methylation is an epigenetic mechanism regulating patterns of gene expression. Our goal was to see if the assessment of DNA methylation might be a useful tool, when used in conjunction with initial, basic in vitro tests, to provide a more informative preliminary appraisal of the toxic potential of chemicals to prioritize them for further evaluation. We sought to give better indications of a compound's toxic potential and its possible mechanism of action at an earlier time and, thereby, contribute to a rational approach of an overall reduction in testing by making improved early decisions. Global and GC-rich patterns of DNA methylation were evaluated along with more traditional cytolethality measurements, e.g., cytolethality and genotoxicity assessments, on rat hepatoma (H4IIE) cells. The relative toxic potential of model compounds camptothecin, 5-fluorouracil, rotenone, and staurosporine was estimated by employing DNA methylation assessments combined with our cytolethality data plus genotoxicity information gleaned from the literature. The overall contribution of the methylation assessment was threefold; it (1) strengthened a ranking based on genotoxicity; (2) provided an indication that a compound might be more potentially problematic than what cytolethality and genotoxicity assessments alone would indicate; and (3) suggested that compounds, particularly nongenotoxins, that are more potent regarding their ability to alter methylation, especially at noncytolethal concentrations, may be more potentially toxic. Altered methylation per se is not proof of toxicity; this needs to be viewed as a component of an evaluation.
Watson
Rebecca E
RE
Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Michigan 48824, USA.
McKim
James M
JM
Cockerell
Gary L
GL
Goodman
Jay I
JI
D000818:Animals; D001374:Azacitidine / Q000378:metabolism / Q000494:pharmacology; D001482:Base Composition / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D003596:Cytosine / Q000737:chemistry / Q000502:physiology; D019175:DNA Methylation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005472:Fluorouracil / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics; D006147:Guanine / Q000737:chemistry / Q000502:physiology; D009152:Mutagenicity Tests / Q000145:classification / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D012402:Rotenone / Q000378:metabolism / Q000494:pharmacology; D019311:Staurosporine / Q000378:metabolism / Q000494:pharmacology
D012402:Rotenone; D006147:Guanine; D003596:Cytosine; D019311:Staurosporine; D001374:Azacitidine; D005472:Fluorouracil