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- Title
INVESTIGATING PROMOTER HYPERMETHYLATION OF APOPTOTIC GENES IN PROSTATE CANCER.
- Authors
Murphy, T. M.; Perry, A. S.; O'Connor, L.; Lawler, M.
- Abstract
It is now well established that cancer cells exhibit a number of genetic defects in the machinery that governs programmed cell death and that sabotage of apoptosis is one of the principal factors aiding in the evolution of the carcinogenic phenotype. A number of studies have implicated aberrant DNA methylation as a key survival mechanism in cancer, whereby promoter hypermethylation silences genes essential for many processes including apoptosis. To date, studies on the methylation profile of apoptotic genes have largely focused on cancers of the breast, colon and stomach, with only limited data available on prostate cancer. The aim of this study was to profile methylation of apoptotic-related genes in order to generate a prostate cancer "apoptotic methylation signature". This in turn could play a role in the early detection and prognosis of prostate cancer and may help elucidate novel therapeutic targets. An in silico approach was first applied to generate a list of apoptotic genes. Relevant genes were identified based on the following criteria: 1) biological role in apoptosis, 2) the presence of a 5' CpG Island 3) susceptibility to promoter hypermethylation in other cancer types and 4) down-regulation in prostate cancer. Under these criteria, 22 apoptoticrelated genes were identified as possible targets of methylation in prostate cancer. PCR assays were designed to amplify whole CpG islands in these gene promoters. Genes will be screened for CpG methylation in a panel of prostate cancer cell lines (LNCaP, DU145, PC-3, 22RV1, RC58) using an automated Denaturing High Performance Liquid Chromatography (DHPLC) instrument (WAVE®, Transgenomic Inc). To date, DHPLC results suggest that the CpG promoter region of TMS1, C-FLIP and BNIP3 are fully or partially methylated in the five cell lines examined, while APAF1, CASP8 and CASP3 show no evidence of CpG promoter methylation. Currently we are screening BIK, DR4 and DR5 for promoter hypermethylation. Genes of interest will be further validated through bisulfite sequencing and methylation levels quantified using quantitative methylation specific PCR in a prostate cancer biorepository that we have generated in Ireland, representing prostate cancer, normal adjacent prostate and benign prostatic hyperplasia.
- Subjects
IRELAND; CANCER cells; CELL death; PHENOTYPES; DNA; METHYLATION; PROSTATE cancer
- Publication
Ulster Medical Journal, 2009, Vol 78, Issue 1, p67
- ISSN
0041-6193
- Publication type
Article