title: No role for Epstein-Barr virus in Dutch hepatocellular carcinoma: a study at the DNA, RNA and protein levels.
abstract: Epstein-Barr virus (EBV) has been suggested to play a role in hepatocellular carcinoma (HCC). However, reports on detailed EBV transcript analyses in HCCs are limited. It was shown recently that expression of the transforming BARF1 (BamHI A rightward open reading frame 1) gene of EBV is restricted to latently EBV-infected epithelial malignancies, i.e. nasopharyngeal carcinoma and gastric carcinoma. The aim of this study was to test the presence of EBV in Dutch HCCs. A semiquantitative DNA PCR-enzyme immunoassay (PCR-EIA) for the BamHI W fragment of EBV was used to assess the presence of EBV in frozen and paraffin-embedded tissues of 16 HCCs. In addition, several RNA detection techniques, i.e. nucleic acid sequence-based amplification (NASBA), RT-PCR, RNA in situ hybridization (RISH) and immunohistochemistry (IHC), were applied. Five of 16 HCCs and two of four hepatitis C virus hepatitis samples were weakly positive for EBV DNA by PCR-EIA. Using sensitive RNA transcription techniques, no transcripts were found for BARF1, EBNA-1 and BARTs (BamHI A rightward transcripts) in any of the liver tissues tested. In addition, RISH for EBER1/2 and BARTs and IHC for EBNA-1, LMP-1 and ZEBRA, performed on the paraffin-embedded tissue of the PCR-EIA-positive cases and on adjacent non-neoplastic liver tissues, were negative. The absence of epithelial-specific BARF1 transcripts and other EBV transcripts and proteins in the EBV DNA PCR-positive cases argues strongly against a role for EBV in HCC.
authors.affiliation: Department of Pathology, Section Molecular Pathology, Vrije Universiteit Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands.
authors.forename: Axel
authors.initials: A
authors.lastname: Zur Hausen
authors.affiliation: Department of Pathology, Section Molecular Pathology, Vrije Universiteit Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands.
authors.forename: Josine
authors.initials: J
authors.lastname: van Beek
authors.affiliation: Department of Pathology, Section Molecular Pathology, Vrije Universiteit Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Bloemena
authors.affiliation: Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Fiebo J
authors.initials: FJ
authors.lastname: Ten Kate
authors.affiliation: Department of Pathology, Section Molecular Pathology, Vrije Universiteit Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands.
authors.forename: Chris J L M
authors.initials: CJLM
authors.lastname: Meijer
authors.affiliation: Department of Pathology, Section Molecular Pathology, Vrije Universiteit Medical Center, PO Box 7057, 1007 MB Amsterdam, The Netherlands.
authors.forename: Adriaan J C
authors.initials: AJC
authors.lastname: van den Brule
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D020031:Epstein-Barr Virus Infections / Q000821:virology*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism / Q000821:virology; D008113:Liver Neoplasms / Q000821:virology*; D009426:Netherlands; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021161:Self-Sustained Sequence Replication; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins; D015245:Deoxyribonuclease BamHI