title: Methylomics analysis identifies epigenetically silenced genes and implies an activation of β-catenin signaling in cervical cancer.
abstract: Using DNA methylation biomarkers in cancer detection is a potential direction in clinical testing. Some methylated genes have been proposed for cervical cancer detection; however, more reliable methylation markers are needed. To identify new hypermethylated genes in the discovery phase, we compared the methylome between a pool of DNA from normal cervical epithelium (n = 19) and a pool of DNA from cervical cancer tissues (n = 38) using a methylation bead array. We integrated the differentially methylated genes with public gene expression databases, which resulted in 91 candidate genes. Based on gene expression after demethylation treatment in cell lines, we confirmed 61 genes for further validation. In the validation phase, quantitative MSP and bisulfite pyrosequencing were used to examine their methylation level in an independent set of clinical samples. Fourteen genes, including ADRA1D, AJAP1, COL6A2, EDN3, EPO, HS3ST2, MAGI2, POU4F3, PTGDR, SOX8, SOX17, ST6GAL2, SYT9, and ZNF614, were significantly hypermethylated in CIN3+ lesions. The sensitivity, specificity, and accuracy of POU4F3 for detecting CIN3+ lesions were 0.88, 0.82, and 0.85, respectively. A bioinformatics function analysis revealed that AJAP1, EDN3, EPO, MAGI2, and SOX17 were potentially implicated in β-catenin signaling, suggesting the epigenetic dysregulation of this signaling pathway during cervical cancer development. The concurrent methylation of multiple genes in cancers and in subsets of precancerous lesions suggests the presence of a driver of methylation phenotype in cervical carcinogenesis. Further validation of these new genes as biomarkers for cervical cancer screening in a larger population-based study is warranted.
authors: Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan; Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
authors: Yu-Chih
authors: YC
authors: Chen
authors: Rui-Lan
authors: RL
authors: Huang
authors: Yung-Kai
authors: YK
authors: Huang
authors: Yu-Ping
authors: YP
authors: Liao
authors: Po-Hsuan
authors: PH
authors: Su
authors: Hui-Chen
authors: HC
authors: Wang
authors: Cheng-Chang
authors: CC
authors: Chang
authors: Ya-Wen
authors: YW
authors: Lin
authors: Mu-Hsien
authors: MH
authors: Yu
authors: Tang-Yuan
authors: TY
authors: Chu
authors: Hung-Cheng
authors: HC
authors: Lai
mesh_terms: D014408:Biomarkers, Tumor; D063646:Carcinogenesis / Q000235:genetics*; D018899:CpG Islands; D019175:DNA Methylation / Q000235:genetics*; D055088:Early Detection of Cancer; D044127:Epigenesis, Genetic; D005260:Female; D006801:Humans; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology; D051176:beta Catenin / Q000235:genetics*
chemical_list: D014408:Biomarkers, Tumor; D051176:beta Catenin
keywords: CIN; DNA methylation; biomarkers; cervical cancer