Discovery of DNA Hypermethylation Using a DHPLC Screening Strategy

Antoinette S. Perry, Hema Liyanage, Mark Lawler and Karen Woodson

volume 2 | issue 1

january/february/march 2007
Pages: 43 - 49

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Promoter hypermethylation is recognized as a hallmark of human cancer, in addition to conventional mechanisms of gene inactivation. As such, many new technologies have been developed over the past two decades to uncover novel targets of methylation and decipher complex epigenetic patterns. However, many of these are either labour intensive or provide limited data, confined to oligonucleotide hybridization sequences or enzyme cleavage sites and cannot be easily applied to screening large sets of sequences or samples. We present an application of denaturing high performance liquid chromatography (DHPLC), which relies on bisulfite modification of genomic DNA, for methylation screening. We validated DHPLC as a methylation screening tool using GSTP1, a well known target of methylation in prostate cancer. We developed an in silico approach to identify potential targets of promoter hypermethylation in prostate cancer. Using DHPLC, we screened two of these targets LGALS3 and SMAD4 for methylation. We show that DHPLC has an application as a fast, sensitive, quantitative and cost effective method for screening novel targets or DNA samples for DNA methylation.

Authors

Antoinette S. Perry

Trinity College Dublin, Ireland

Hema Liyanage

Transgenomic Inc., Cambridge, MA

Mark Lawler

Trinity College Dublin, Ireland

Karen Woodson

National Cancer Institute, Bethesda, Maryland

We now provide open access to journal articles published online for one year or more. This article may be downloaded at the following link: