and in the first exon, and we demonstrated here that methylation of this CpG is responsible for the repression of DLEC1 expression in uterine leiomyoma. Our analysis revealed a strong association between silencing of DLEC1 expression and promoter hypermethylation in uterine leiomyoma; in addition, treatment of addition “2859375 of cultured primary uterine buy GLYX-13 leiomyoma smooth muscle cells with a DNMT inhibitor restored DLEC1 expression. The DLEC1 gene encodes a 166 kDa protein, whose biologic function remains unknown due to lack of homology to any known conserved proteins or domains. In the future, we plan to characterize the biological function of DLEC1 in uterine leiomyoma. KRT19 is an intermediate filament protein responsible for the structural integrity of epithelial cells, this genes encodes a 40-kDa protein. In mammalian cells, keratin filaments are organized in a complex network spreading from the nucleus to the cytoplasmic membrane. KRT19 is also known as an epigenetically regulated tumor suppressor gene, which has frequently demonstrated promoter hypermethylation associated with transcriptional downregulation in several cancerous tumors such as neuroblastomas, squamous cell carcinoma of the head and neck region and 
renal cell carcinomas. Also, it is one of the most common used markers for real-time RT-PCR detection of tumor cells disseminated in lymph nodes, peripheral blood and bone marrow of breast cancer patients. Using genome-wide analyses of DNA methylation in 12829792 uterine leiomyoma we hope to define a specific epigenetic profile that could inform the development of diagnostic biomarkers for uterine leiomyoma as well as identify potential therapeutic targets. Because DNA methylation is reversible, epigenetic modifying drugs could be used in the medical management of uterine leiomyoma. Importantly, aberrant DNA methylation and other epigenetic abnormalities may represent a critical initial mechanism that triggers transformation of a single myometrial cell that will eventually give rise to a monoclonal leiomyoma tumor. Understanding the mechanism underlying the pathogenesis of uterine leiomyoma will be critical for developing new preventive and therapeutic approaches to the disease. Materials and Methods Ethics Statement To obtain human tissues, we followed the protocol approved by the Institutional Review Board for Human Research of Northwestern University and New York University. Written informed consent was received from all subjects. Tissue acquisition For in vivo studies, we obtained matched pairs of leiomyoma and adjacent myometrium from a total of 23 African American and 14 Caucasian-American subjects undergoing hysterectomy for symptomatic fibroids. To minimize heterogeneity due to race we used samples from 18 African American subjects for both genome-wide DNA methylation and gene expression microarrays. In follow-up verification studies, we included samples from 4 of the original African American group plus 4 additional Caucasian subjects for bisulfite sequencing and all 18 original African American plus 10 Caucasian subjects for mRNA quantification using real-time RTPCR. Samples from Caucasian subjects were added to evaluate whether similar patterns of DNA methylation and mRNA expression were observed. Key clinical characteristics of the 18 African American subjects, whose samples were used for both microarrays are described in Primary cell isolation Leiomyoma smooth muscle cells were isolated from the peripheral portions approximat
