Molecular mechanisms of aberrant DNA methylation during multistage carcinogenesis

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The aim of this study is to detect the molecules which associate with DNA methyltransferase (DNMT)1 and contribute to induce aberrant DNA methylation profiles during human carcinogenesis. We used HepG2 cell line which was derived from human liver cancer and showed high expression levels of DNMT1. We performed immunoprecipitation of DNMT1 using optimum antibody in HepG2 nuclear extract, and separated immunoprecipitant by SDS-PAGE. Co-precipitated proteins were analyzed by 2 Dimensional Image Converted Analysis of LCMS (2D-ICAL method), and the candidate molecule which may associate DNMT1 and may contribute aberrant DNA methylation was identified. Direct association between the candidate molecule and DNMT1 is being evaluated. Mechanisms how the candidate molecule participates in aberrant distribution of DNMT1 resulting in aberrant DNA methylation profiles will be examined.