title: Genome-wide methylation profiling identifies hypermethylated biomarkers in high-grade cervical intraepithelial neoplasia.
abstract: Epigenetic modifications, such as aberrant DNA promoter methylation, are frequently observed in cervical cancer. Identification of hypermethylated regions allowing discrimination between normal cervical epithelium and high-grade cervical intraepithelial neoplasia (CIN2/3), or worse, may improve current cervical cancer population-based screening programs. In this study, the DNA methylome of high-grade CIN lesions was studied using genome-wide DNA methylation screening to identify potential biomarkers for early diagnosis of cervical neoplasia. Methylated DNA Immunoprecipitation (MeDIP) combined with DNA microarray was used to compare DNA methylation profiles of epithelial cells derived from high-grade CIN lesions with normal cervical epithelium. Hypermethylated differentially methylated regions (DMRs) were identified. Validation of nine selected DMRs using BSP and MSP in cervical tissue revealed methylation in 63.2-94.7% high-grade CIN and in 59.3-100% cervical carcinomas. QMSP for the two most significant high-grade CIN-specific methylation markers was conducted exploring test performance in a large series of cervical scrapings. Frequency and relative level of methylation were significantly different between normal and cancer samples. Clinical validation of both markers in cervical scrapings from patients with an abnormal cervical smear confirmed that frequency and relative level of methylation were related with increasing severity of the underlying CIN lesion and that ROC analysis was discriminative. These markers represent the COL25A1 and KATNAL2 and their observed increased methylation upon progression could intimate the regulatory role in carcinogenesis. In conclusion, our newly identified hypermethylated DMRs represent specific DNA methylation patterns in high-grade CIN lesions and are candidate biomarkers for early detection.
authors.affiliation: Department of Gynaecological Oncology, University of Groningen, University Medical Centre Groningen, Groningen, The Netherlands.
authors.forename: Ágnes
authors.initials: Á
authors.lastname: Lendvai
authors.forename: Frank
authors.initials: F
authors.lastname: Johannes
authors.forename: Christina
authors.initials: C
authors.lastname: Grimm
authors.forename: Jasper J H
authors.initials: JJ
authors.lastname: Eijsink
authors.forename: René
authors.initials: R
authors.lastname: Wardenaar
authors.forename: Haukeline H
authors.initials: HH
authors.lastname: Volders
authors.forename: Harry G
authors.initials: HG
authors.lastname: Klip
authors.forename: Harry
authors.initials: H
authors.lastname: Hollema
authors.forename: Ritsert C
authors.initials: RC
authors.lastname: Jansen
authors.forename: Ed
authors.initials: E
authors.lastname: Schuuring
authors.forename: G Bea A
authors.initials: GB
authors.lastname: Wisman
authors.forename: Ate G J
authors.initials: AG
authors.lastname: van der Zee
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics*; D016022:Case-Control Studies; D019175:DNA Methylation; D005260:Female; D015894:Genome, Human; D006801:Humans; D000074201:Katanin; D008875:Middle Aged; D024041:Non-Fibrillar Collagens / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D017422:Sequence Analysis, DNA; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis / Q000235:genetics* / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; C456021:COL25A1 protein, human; D024041:Non-Fibrillar Collagens; D000251:Adenosine Triphosphatases; C573101:KATNAL2 protein, human; D000074201:Katanin