Methylated Host Cell Gene Promoters and Human Papillomavirus Type 16 and 18 Predicting Cervical Lesions and Cancer

PLoS One. 2015 Jun 9;10(6):e0129452. doi: 10.1371/journal.pone.0129452. eCollection 2015.

Abstract

Change in the host and/or human papillomavirus (HPV) DNA methylation profile is probably one of the main factors responsible for the malignant progression of cervical lesions to cancer. To investigate those changes we studied 173 cervical samples with different grades of cervical lesion, from normal to cervical cancer. The methylation status of nine cellular gene promoters, CCNA1, CDH1, C13ORF18, DAPK1, HIC1, RARβ2, hTERT1, hTERT2 and TWIST1, was investigated by Methylation Specific Polymerase Chain Reaction (MSP). The methylation of HPV18 L1-gene was also investigated by MSP, while the methylated cytosines within four regions, L1, 5'LCR, enhancer, and promoter of the HPV16 genome covering 19 CpG sites were evaluated by bisulfite sequencing. Statistically significant methylation biomarkers distinguishing between cervical precursor lesions from normal cervix were primarily C13ORF18 and secondly CCNA1, and those distinguishing cervical cancer from normal or cervical precursor lesions were CCNA1, C13ORF18, hTERT1, hTERT2 and TWIST1. In addition, the methylation analysis of individual CpG sites of the HPV16 genome in different sample groups, notably the 7455 and 7694 sites, proved to be more important than the overall methylation frequency. The majority of HPV18 positive samples contained both methylated and unmethylated L1 gene, and samples with L1-gene methylated forms alone had better prognosis when correlated with the host cell gene promoters' methylation profiles. In conclusion, both cellular and viral methylation biomarkers should be used for monitoring cervical lesion progression to prevent invasive cervical cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Biomarkers / metabolism
  • Cell Line
  • Cervix Uteri / metabolism
  • Cervix Uteri / pathology
  • CpG Islands / genetics
  • Cytosine / metabolism
  • DNA Methylation / genetics*
  • DNA, Viral / genetics
  • Female
  • Gene Expression Regulation, Viral / genetics
  • Human papillomavirus 16 / genetics*
  • Human papillomavirus 18 / genetics*
  • Humans
  • Middle Aged
  • Papillomavirus Infections / genetics*
  • Papillomavirus Infections / pathology
  • Promoter Regions, Genetic / genetics*
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / pathology
  • Uterine Cervical Neoplasms / virology*
  • Young Adult

Substances

  • Biomarkers
  • DNA, Viral
  • Cytosine

Grant support

This work was supported by the Croatian Ministry of Science, Education and Sport (grant 098-0982464-2510). MG was also supported by the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement No 316289. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.