Detection of methylation status of Epstein-Barr virus DNA C promoter in the diagnosis of nasopharyngeal carcinoma

Cancer Sci. 2020 Feb;111(2):592-600. doi: 10.1111/cas.14281. Epub 2020 Feb 5.

Abstract

The detection of Epstein-Barr virus (EBV) DNA load in nasopharyngeal (NP) brushing samples for diagnosis of nasopharyngeal carcinoma (NPC) has attracted great attention. Further improvements that eliminate the need for clinical settings will greatly extend its application. A total of 250 participants were recruited to obtain NP brushing samples. Brush sampling both with and without the guide of endoscopy was conducted in 38 NPC patients. EBV DNA load, EBV RNA transcript and EBV DNA C promoter methylation status were, respectively, evaluated. Typical latency II transcripts were observed in brushing samples from NPC patients but not controls. Unlike in tissues, multiple lytic gene transcripts were observed not only in NPC patients but also in controls. Apart from EBV RNA transcript, samples from NPC patients also showed higher levels of EBV DNA load and C promoter methylation degree than their controls. Qualitative analysis further showed that EBV DNA C promoter was methylated in all NPC patients but in only 18.4% of the control group. Combined analysis of EBV DNA methylated degree and EBV DNA load increased the sensitivity to 100% in the detection of NPC. Using qualitative methylated type as the criteria, up to 89.5% of samples collected via blind brushing showed consistent results with samples collected via endoscopy-guided brushing from NPC patients. Detection of the methylation status of EBV DNA C promoter in NP brushing samples shows great potential in diagnosing NPC and may provide an appealing alternative for the non-invasive detection and screening of NPC without the need for clinical settings.

Keywords: Epstein-Barr virus DNA load; Epstein-Barr virus DNA methylation; diagnosis; nasopharyngeal brushing; nasopharyngeal carcinoma.

MeSH terms

  • DNA Methylation*
  • DNA, Viral / genetics
  • Epstein-Barr Virus Infections / complications
  • Epstein-Barr Virus Infections / genetics*
  • Female
  • Gene Expression Regulation, Viral
  • Herpesvirus 4, Human / genetics*
  • Herpesvirus 4, Human / physiology
  • Humans
  • Male
  • Nasopharyngeal Carcinoma / diagnosis*
  • Nasopharyngeal Carcinoma / virology
  • Nasopharyngeal Neoplasms / diagnosis*
  • Nasopharyngeal Neoplasms / virology
  • Promoter Regions, Genetic
  • Sensitivity and Specificity
  • Viral Proteins / genetics
  • Virus Latency

Substances

  • DNA, Viral
  • Viral Proteins