Hepatitis C and B virus infections in hepatocellular carcinoma. Analysis of direct detection of viral genome in paraffin embedded tissues

Cancer. 1996 May 1;77(9):1787-91. doi: 10.1002/(SICI)1097-0142(19960501)77:9<1787::AID-CNCR5>3.0.CO;2-9.


Background: Although there have been many seroepidemiologic studies on hepatitis C virus (HCV) infection and hepatocellular carcinoma (HCC) occurrence, the actual role of HCV in hepatocarcinogenesis is unknown.

Methods: We have previously reported on a highly sensitive method of detecting and identifying sequences of RNA genome in formalin fixed, paraffin embedded (FFPE) tissue by polymerase chain reaction (PCR) assay. Using this method, we carried out a retrospective study to determine the prevalence of HCV and hepatitis B virus (HBV) genomes in FFPE specimens from 102 Japanese patients with HCC.

Results: HCV-RNA was detected by nested PCR reverse transcription (RT) in 64 of the 102 patients (62.7%), and 78.1% (50/64) of those HCV-RNA-positive patients had HCV genotype II. HCV-RNA was present in 54 of 70 (77.1%) anti-HCV-positive patients, and also in 5 of 20 (25%) anti-HCV-negative patients. HBV-DNA was detected by nested PCR in FFPE liver specimens from 21 of 102 (20.6%) patients. HBV-DNA positivity was consistent with seropositivity for serum HBV markers in 17 of these 21 patients (80.9%). HBV-DNA was present in FFPE samples from 2 patients who were seronegative for HBV markers, and in 1 patient who was seropositive for anti-HBs. Double infection of these two viruses was found in 6 patients (5.9%). Three patients (2.9%) were negative for both hepatic viral genomes and serum viral markers.

Conclusions: The precise prevalence of HCV and/or HBV infection among HCC patients can be determined by studying routinely-processed FFPE HCC samples preserved for up to 11 years using the technique of nested PCR. HCV-RNA was detected in the majority of our HCC cases; type II was the most common genotype of HCV encountered. The incidence of HCV-associated HCC was three times greater than that of HBV. Thus, the hepatitis virus infection most frequently associated, and probably ectologically implicated, with HCC in Japanese people is HCV infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Carcinoma, Hepatocellular / complications*
  • DNA, Viral / analysis
  • DNA, Viral / genetics
  • Female
  • Fixatives
  • Formaldehyde
  • Genome, Viral*
  • Genotype
  • Hepacivirus / genetics*
  • Hepatitis B / complications*
  • Hepatitis B Antibodies / blood
  • Hepatitis B virus / genetics*
  • Hepatitis C / complications*
  • Hepatitis C Antibodies / blood
  • Humans
  • Japan
  • Liver Neoplasms / complications*
  • Male
  • Middle Aged
  • Paraffin Embedding
  • Polymerase Chain Reaction
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • Retrospective Studies
  • Seroepidemiologic Studies
  • Transcription, Genetic


  • DNA, Viral
  • Fixatives
  • Hepatitis B Antibodies
  • Hepatitis C Antibodies
  • RNA, Viral
  • Formaldehyde