title: Human Papillomavirus Type 16 L2 DNA Methylation in Exfoliated Cervical Cells From College-Age Women.
abstract: The Carolina Women's Care Study (CWCS) at the University of South Carolina followed 467 young women with the goal of identifying biomarkers of human papillomavirus (HPV) persistence. In this study, we analyzed the methylation of HPV16 DNA.
  
  
  
  The aims of this study were to determine the methylation status of the HPV16 L2 gene in DNA isolated from exfoliated cervical cells collected longitudinally as part of the CWCS and to determine the prevalence of polymorphisms (single nucleotide polymorphisms [SNPs]) in folate metabolizing enzymes and DNA repair enzymes known to affect DNA methylation in blood-derived genomic DNA from CWCS participants. For methylation studies, DNA samples were bisulfite converted and amplified with the EpiTect Whole Bisulfitome kit. Polymerase chain reaction was performed for amplicons containing 5 CpG sites in L2. Pyrosequencing was carried out using EpigenDx and analyzed with PyroMark Software. Taqman genotyping assays were performed to determine selected SNP alleles in the CWCS cohort.
  
  
  
  Methylation data were obtained for 82 samples from 27 participants. Of these, 22 participants were positive for HPV16 for 3 or more visits (≥12 months). Methylation in L2 was detectable, but methylation levels varied and were not associated with HPV16 persistence. No linearity of methylation levels over time was observed in participants for whom longitudinal data could be analyzed. Analysis of 9 selected SNPs did not reveal an association with persistence. We conclude that at early stages of infection methylation of HPV16 L2 DNA in Pap test samples is not a predictive biomarker of HPV persistence.
authors: 1Department of Drug Discovery and Biomedical Sciences, South Carolina College of Pharmacy, University of South Carolina, Columbia, South Carolina; 2Department of Pharmaceutical Sciences, College of Pharmacy, Presbyterian College, Clinton, South Carolina; and 3Department of Pathology, Microbiology and Immunology, School of Medicine, University of South Carolina, Columbia, South Carolina.
authors: Erin L
authors: EL
authors: Anderson
authors: Carolyn E
authors: CE
authors: Banister
authors: Susannah
authors: S
authors: Kassler
authors: Amy
authors: A
authors: Messersmith
authors: Lucia
authors: L
authors: Pirisi
authors: Kim E
authors: KE
authors: Creek
authors: Michael D
authors: MD
authors: Wyatt
mesh_terms: D000328:Adult; D036022:Capsid Proteins / Q000235:genetics*; D002584:Cervix Uteri / Q000821:virology*; D019175:DNA Methylation; D045643:DNA Repair Enzymes / Q000235:genetics; D004279:DNA, Viral / Q000378:metabolism*; D004847:Epithelial Cells / Q000821:virology*; D005260:Female; D006801:Humans; D008137:Longitudinal Studies; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D011446:Prospective Studies; D017422:Sequence Analysis, DNA; D013022:South Carolina; D013334:Students; D055815:Young Adult
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; C063316:L2 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D045643:DNA Repair Enzymes