Quantification of hepatitis B virus covalently closed circular DNA in patients with hepatocellular carcinoma

J Hepatol. 2006 Oct;45(4):553-9. doi: 10.1016/j.jhep.2006.05.014. Epub 2006 Jun 23.

Abstract

Background/aims: This study aimed to measure the intrahepatic total hepatitis B virus (HBV) DNA and covalently closed circular DNA (cccDNA) levels in tumor and non-tumor tissues in hepatocellular carcinoma (HCC) patients.

Methods: Intrahepatic total HBV DNA and cccDNA were measured in 25 HCC patients (21 hepatitis B surface antigen [HBsAg]-positive and 4 HBsAg-negative) by the Invader assay.

Results: A low level of intrahepatic HBV DNA was detectable in all HBsAg-negative patients. For HBsAg-positive patients, the intrahepatic total HBV DNA levels in the tumor and non-tumor tissues were comparable (P=0.903). However, the tumor tissues had significantly higher levels of cccDNA (0.35 vs. 0.16 copies/cell, P=0.030) and higher proportion of intrahepatic HBV DNA in the form of cccDNA (100% vs. 84%, P=0.004) than the non-tumor tissues. Seventeen out of 21 (81%) tumor tissues had intrahepatic HBV DNA solely in cccDNA form. Analysis of HBV mRNA expression indicated that HBV replication appeared to be lower in the tumor tissues than the non-tumor tissues.

Conclusions: Compared to the non-tumor tissues, the levels of HBV replication in the tumor tissues appeared to be lower, and cccDNA was the predominant form of HBV DNA in the tumor tissues.

MeSH terms

  • Adult
  • Aged
  • Carcinoma, Hepatocellular / virology*
  • DNA, Circular / analysis
  • DNA, Circular / isolation & purification*
  • DNA, Viral / analysis
  • DNA, Viral / isolation & purification
  • Female
  • Gene Expression Regulation, Viral
  • Hepatitis B Surface Antigens / metabolism
  • Hepatitis B virus / genetics*
  • Hepatitis B, Chronic / complications
  • Hepatitis B, Chronic / virology*
  • Humans
  • Liver / virology
  • Liver Neoplasms / virology*
  • Male
  • Middle Aged
  • RNA, Messenger / analysis
  • RNA, Messenger / isolation & purification
  • RNA, Viral / analysis
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • DNA, Circular
  • DNA, Viral
  • Hepatitis B Surface Antigens
  • RNA, Messenger
  • RNA, Viral