title: DNA methylation analysis in liquid-based cytology for cervical cancer screening.
abstract: Cervical cancer is the second most common type of cancer in women worldwide. Preinvasive disease can be detected by cervical cytology. All currently available cytology technologies rely on the visual analysis of exfoliated cells from the uterine cervix. Improvement of conventional cytological screening has been proposed by the introduction of molecular-based markers applied to liquid-based cytology (LBC), the suspension of cells collected from the cervix. DNA methylation changes occur very early in carcinogenesis and identification of appropriate DNA methylation markers in such samples should be able to distinguish high-grade squamous intraepithelial lesions (HSIL) from nonspecific cytology changes and the normal cervix. To address this potential, we have undertaken a proof-of-principle study of methylation status of LBC samples from HSIL cytology cases compared against matched normal controls. Using quantitative methylation-specific PCR on 28 genes, we found SOX1, HOXA11 and CADM1 to significantly discriminate between the groups analyzed (p<0.01). Area under the receiver operating characteristic (ROC) curve (AUC) demonstrated that methylation of SOX1, HOXA11 and CADM1 could discriminate between HSIL cases and controls with high sensitivity and specificity (AUC 0.910, 0.844 and 0.760, respectively). The results were further validated in an independent set. This proof-of-principle study is the first to validate the results in an independent case/control set and presents HOXA11, a gene that is important for cervical development, as a potentially useful DNA marker in LBC samples. Further assessment of these preliminary estimates will need to be performed in a larger cohort to confirm clinical utility.
authors.affiliation: Department of Gynecological Oncology, Institute for Women's Health, University College London, London WC1E 6DH, United Kingdom.
authors.forename: Sophia
authors.initials: S
authors.lastname: Apostolidou
authors.forename: Richard
authors.initials: R
authors.lastname: Hadwin
authors.forename: Matthew
authors.initials: M
authors.lastname: Burnell
authors.forename: Allison
authors.initials: A
authors.lastname: Jones
authors.forename: Donna
authors.initials: D
authors.lastname: Baff
authors.forename: Nitisha
authors.initials: N
authors.lastname: Pyndiah
authors.forename: Tim
authors.initials: T
authors.lastname: Mould
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Jacobs
authors.forename: Simon
authors.initials: S
authors.lastname: Beddows
authors.forename: Gabrijela
authors.initials: G
authors.lastname: Kocjan
authors.forename: Martin
authors.initials: M
authors.lastname: Widschwendter
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D016022:Case-Control Studies; D002584:Cervix Uteri / Q000473:pathology*; D003584:Cytological Techniques; D019175:DNA Methylation; D005260:Female; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D008403:Mass Screening; D008875:Middle Aged; D009367:Neoplasm Staging; D027383:Papillomaviridae / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011379:Prognosis; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000235:genetics / Q000821:virology
chemical_list: D014408:Biomarkers, Tumor; C484298:HOXA11 protein, human; D018398:Homeodomain Proteins