title: L1 gene methylation in high-risk human papillomaviruses for the prognosis of cervical intraepithelial neoplasia.
abstract: OBJECTIVE
  The purpose of this study was to investigate the clinical significance of DNA methylation of the human papillomavirus (HPV) genome as a prognostic biomarker for cervical intraepithelial neoplasia (CIN).
  
  
  METHODS
  Clinical samples (paraffin-embedded tissues obtained by conization/hysterectomy or initial punch biopsy) were collected from patients at the Gynecologic Oncology of the Hyogo Cancer Center with informed consent. We evaluated the methylation status of the L1 gene of the HPV genome by bisulfite sequencing, calculating the methylation ratio (L1MR) as (number of methylated CpGs in the analyzed region of the L1 gene) / (number of all CpGs in the analyzed region of the L1 gene) × 100. The methylation analysis and in situ hybridization were performed with serial tissue-section slices.
  
  
  RESULTS
  DNA methylation was observed in the L1 gene, but not in the long control region of HPV-16, -18, or the other high-risk HPV types including HPV-31, -52, and -58. L1MR was associated with the CIN grade; the median L1MR was 2.3%, 11.2%, 35.2%, and 50.0% for CIN1, CIN2, CIN3, and squamous cell carcinoma, respectively. L1MRs also seemed to indicate physical status (integrated or episomal form) of the HPV genome in the host cell. L1MR of the progression group was significantly higher than that of the regression group.
  
  
  CONCLUSIONS
  L1MR was associated with the CIN grade and indicated the HPV genome status in the host cell: high L1MR indicated HPV genome integration linked to progression from early-stage CINs, whereas low L1MR indicated an episomal HPV genome location in host cells. L1MR may be a prognostic indicator of CIN.
authors: Section of Translational Research, Hyogo Cancer Center, Akashi, Japan.
authors: Noriko
authors: N
authors: Oka
authors: Masahiro
authors: M
authors: Kajita
authors: Ryuichiro
authors: R
authors: Nishimura
authors: Chiho
authors: C
authors: Ohbayashi
authors: Tamotsu
authors: T
authors: Sudo
mesh_terms: D036022:Capsid Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D018899:CpG Islands; D019175:DNA Methylation / Q000502:physiology; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005260:Female; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D006801:Humans; D017403:In Situ Hybridization; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D030361:Papillomavirus Infections / Q000150:complications / Q000175:diagnosis / Q000235:genetics / Q000821:virology; D016133:Polymerase Chain Reaction; D011379:Prognosis; D012307:Risk Factors; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000209:etiology / Q000821:virology; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000209:etiology / Q000821:virology
chemical_list: D036022:Capsid Proteins; D004279:DNA, Viral; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral