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Supplement: Trans-HHS Workshop: Diet, DNA Methylation Processes and Health

Genistein Alters Methylation Patterns in Mice^{1 ,2}

J. Kevin Day^*,,***,3, Andrew M. Bauer^*, Charles desBordes, Yi Zhuang^*, Byung-Eun Kim^*, Leslie G. Newton^*, Vedika Nehra^*, Kara M. Forsee^*, Ruth S. MacDonald^**, Cynthia Besch-Williford, Tim Hui-Ming Huang and Dennis B. Lubahn^*,,#,***,##4

^* Departments of Biochemistry, Pathology, Child Health, ^# Animal Sciences, ^** Nutrition and Veterinary Pathobiology, ^*** Genetics and ^## Molecular Biology Programs, University of Missouri, Columbia, MO 65211, and Department of Biology, City University of New York, NY 11225

⁴To whom correspondence should be addressed. E-mail: lubahnd{at}missouri.edu.

In this study we examine the effect of the phytoestrogen genistein on DNA methylation. DNA methylation is thought to inhibit transcription of genes by regulating alterations in chromatin structure. Estrogenic compounds have been reported to regulate DNA methylation in a small number of studies. Additionally, phytoestrogens are believed to affect progression of some human diseases, such as estrogen-dependent cancers, osteoporosis and cardiovascular disease. Specifically, our working hypothesis is that certain soy phytoestrogens, such as genistein, may be involved in preventing the development of certain prostate and mammary cancers by maintaining a protective DNA methylation profile. The objective of the present study is to use mouse differential methylation hybridization (DMH) arrays to test for changes in the methylation status of the cytosine guanine dinucleotide (CpG) islands in the mouse genome by examining how these methylation patterns are affected by genistein. Male mice were fed a casein-based diet (control) or the same diet containing 300 mg genistein/kg according to one of four regimens: control diet for 4 wk, genistein diet for 4 wk, control diet for 2 wk followed by genistein diet for 2 wk and genistein diet for 2 wk followed by control diet for 2 wk. DNA from liver, brain and prostate were then screened with DMH arrays. Clones with methylation differences were sequenced and compared with known sequences. In conclusion, consumption of genistein diet was positively correlated with changes in prostate DNA methylation at CpG islands of specific mouse genes.

KEY WORDS: • genistein • phytoestrogen • DNA methylation

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