title: Methylation pattern ofCDH1promoter and its association withCDH1gene expression in cytological cervical specimens.
abstract: Cervical cancer is the fourth leading cause of cancer mortality in females worldwide. Infection with high-risk human papillomavirus (HPV) is essential but insufficient to cause cervical cancer, and the clearance of HPV infection is mediated by the immune system. The deficit of molecules responsible for adhesion may play a role in the development of cervical cancer. E-cadherin is encoded by the cadherin 1 (CDH1) gene, and is involved in cell adhesion by forming adherens junctions. The aim of present study was to investigate the methylation pattern of theCDH1promoter and to identify the association betweenCDH1promoter hypermethylation,CDH1gene expression and HPV infection in cervical specimens obtained from 93 patients with low-grade squamous intraepithelial lesions (SILs), high-grade SILs or squamous cell carcinomas, and from 47 patients with normal cervical cytology (HPV-negative). The methylation pattern of theCDH1promoter was investigated by methylation-specific polymerase chain reaction and quantitative pyrosequencing.CDH1gene expression was measured by relative quantification.CDH1methylation was significantly higher in both types of lesions and in cervical cancer than in normal samples, andCDH1gene expression was significantly reduced during SIL progression (P=0.0162). However, the influence of HPV infection or HPV E6 expression on the methylation pattern of theCDH1gene or its gene expression levels could not be confirmed. The present results support that the methylation of theCDH1gene is age-related in patients with cervical lesions (P=0.01085), and therefore, older patients could be more susceptible to cancer than younger patients. The important methylation of theCDH1promoter occurred near the transcription factor binding sites on nucleotides -13 and +103, which are close to the translational start codon. These results suggest that methylation at these sites may be an important event in the transcriptional regulation of E-cadherin, and in patients harboring these methylated cytosines, this event may facilitate HPV-driven carcinogenesis.
authors.affiliation: Department of Oncology, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Veronika
authors.initials: V
authors.lastname: Holubeková
authors.affiliation: Department of Molecular Medicine, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Andrea
authors.initials: A
authors.lastname: Mendelová
authors.affiliation: Bioinformatic Unit, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Marián
authors.initials: M
authors.lastname: Grendár
authors.affiliation: Department of Oncology, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Sandra
authors.initials: S
authors.lastname: Meršaková
authors.affiliation: Clinic of Gynecology and Obstetrics, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Ivana
authors.initials: I
authors.lastname: Kapustová
authors.affiliation: Department of Oncology, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Karin
authors.initials: K
authors.lastname: Jašek
authors.affiliation: Department of Molecular Biology, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Andrea
authors.initials: A
authors.lastname: Vaňochová
authors.affiliation: Clinic of Gynecology and Obstetrics, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Jan
authors.initials: J
authors.lastname: Danko
authors.affiliation: Department of Oncology, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia; Department of Molecular Medicine, Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin, Biomedical Center Martin, SK-03601 Martin, Slovakia.
authors.forename: Zora
authors.initials: Z
authors.lastname: Lasabová
keywords: CDH1 promoter; cervical cancer; gene expression; methylation; pyrosequencing