co-localization of the oncogenic transcription factor mycn and the dna methyl binding protein mecp2 at genomic sites in neuroblastomaco-localization致癌基因的转录因子mycn和基因组的dna结合蛋白甲酯mecp2网站在神经母细胞瘤.pdfVIP

Co-Localization of the Oncogenic Transcription Factor MYCN and the DNA Methyl Binding Protein MeCP2 at Genomic Sites in Neuroblastoma Derek M. Murphy1,2, Patrick G. Buckley1,2,3, Sudipto Das1,2, Karen M. Watters1,2, Kenneth Bryan1,2, Raymond L. Stallings1,2* 1 Department of Cancer Genetics, Royal College of Surgeons in Ireland, Dublin, Ireland, 2 Children’s Research Centre, Our Lady’s Children’s Hospital, Crumlin, Dublin, Ireland, 3 Department of Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, Dublin, Ireland Abstract Background: MYCN is a transcription factor that is expressed during the development of the neural crest and its dysregulation plays a major role in the pathogenesis of pediatric cancers such as neuroblastoma, medulloblastoma and rhabdomyosarcoma. MeCP2 is a CpG methyl binding protein which has been associated with a number of cancers and developmental disorders, particularly Rett syndrome. Methods and Findings: Using an integrative global genomics approach involving chromatin immunoprecipitation applied to microarrays, we have determined that MYCN and MeCP2 co-localize to gene promoter regions, as well as inter/intragenic sites, within the neuroblastoma genome (MYCN amplified Kelly cells) at high frequency (70.2% of MYCN sites were also positive for MeCP2). Intriguingly, the frequency of co-localization was significantly less at promoter regions exhibiting substantial hypermethylation (8.7%), as determined by methylated DNA immunoprecipitation (MeDIP) applied to the same microarrays. Co-immunoprecipitation of MYCN using an anti-MeCP2 antibody indicated that a MYCN/MeCP2 interaction occurs at protein level. mRNA expression profiling revealed that the median expression of genes with promoters bound by MYCN was significantly higher tha