Methylation target array for rapid analysis of CpG island hypermethylation in multiple tissue genomes

摘要: Hypermethylation of multiple CpG islands is a common event in cancer. To assess the prognostic values this epigenetic alteration, we developed Methylation Target Array (MTA), derived from concept tissue microarray, for simultaneous analysis DNA hypermethylation hundreds genomes. In MTA, linker-ligated island fragments were digested with methylation-sensitive endonucleases and amplified flanking primers. A panel 468 MTA amplicons, which represented whole repertoire methylated 93 breast tumors, 20 normal tissues, 4 cancer cell lines, arrayed on nylon membrane probe hybridization. Positive hybridization signals detected tumor but not indicative aberrant samples. This attributed to sites that protected restriction by PCR samples, while same restricted could be frequencies 10 genes tested tumors lines 60% GPC3, 58% RASSF1A, 32% 3OST3B, 30% HOXA5, 28% uPA, 25% WT1, 23% BRCA1, 9% DAPK1, 0% KL. Furthermore, 5 7 loci these was significantly correlated hormone receptor status, clinical stages, ages at diagnosis patients analyzed. novel approach thus provides an additional avenue assessing clinicopathological consequences