title: Methylation status of the E2 binding sites of HPV16 in cervical lesions determined with the Luminex® xMAP™ system.
abstract: Cervical carcinogenesis is driven by deregulated E6/E7 expression in dividing cells. A potential deregulating mechanism is methylation of E2 binding sites in the viral long control region, thereby prohibiting HPVE2-mediated transcription regulation. Here the frequency of HPV16E2BS methylation in cervical lesions (SCC, n=29; CIN3, n=17) and scrapes (controls, n=17; CIN3, n=21) was investigated. Three E2BSs were amplified using methylation independent PCR followed by specific detection of methylated CpGs using the Luminex® xMAP™ system. The frequency of E2BS1, E2BS3 and E2BS4 methylation was significantly higher in SCC compared to CIN3, i.e. 93% vs. 21% (p<0.01), 90% vs. 47% (p<0.01) and 69% vs. 5% (p<0.01), respectively and ranged from 6 to 15% in controls. In scrapings of women with CIN3 methylation ranged from 24 to 33%. In conclusion, we showed that the MIP-Luminex system is a highly sensitive method for methylation analysis. HPV16 E2BSs methylation appeared highly frequent in SCC, with particularly E2BS3 methylation occurring proportional to severity of cervical disease.
authors.affiliation: Department of Pathology, Unit of Molecular Pathology, VU University Medical Center, 1007 MB, Amsterdam, The Netherlands.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Snellenberg
authors.forename: Denise M
authors.initials: DM
authors.lastname: Schütze
authors.forename: Debbie
authors.initials: D
authors.lastname: Claassen-Kramer
authors.forename: Chris J L M
authors.initials: CJ
authors.lastname: Meijer
authors.forename: Peter J F
authors.initials: PJ
authors.lastname: Snijders
authors.forename: Renske D M
authors.initials: RD
authors.lastname: Steenbergen
mesh_terms: D000328:Adult; D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000821:virology; D002478:Cells, Cultured; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D052162:Human papillomavirus 16 / Q000378:metabolism*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008745:Methylation; D008875:Middle Aged; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000821:virology*; D002578:Uterine Cervical Dysplasia / Q000821:virology
chemical_list: D004268:DNA-Binding Proteins; C054855:E2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral