Detection of integrated hepatitis B virus DNA in hepatocellular carcinoma cell lines by nonradioactive in situ hybridization
- 1 April 1990
- journal article
- research article
- Published by Wiley in Journal of Medical Virology
- Vol. 30 (4) , 266-271
- https://doi.org/10.1002/jmv.1890300407
Abstract
A sensitive and specific nonradioactive in situ hybridization method capable of detecting single‐copy integrated hepatitis B virus (HBV) DNA sequences in hepatocellular carcinoma (HCC) cell lines was developed. In situ hybridization of biotinylated HBV (adr, adw) DNA probes with nine different human HCC cell lines were carried out in 96‐well microtiter plates. Integration was detected in HCC cell lines HCCM, Hep3B, huH‐1, huH‐4, and PLC/PRF/5. Detection of single‐copy HBV DNA sequences was also achieved in Hep3B and huH‐4. HCC cell lines HepG2, HUH‐6, HUH‐7, Mahlavu, and the non‐HCC control MCF‐7, gave clear negative results. These results show a 100% correlation with those obtained by Southern blot hybridization assay. The results demonstrate that nonradioactive detection of single‐copy integrated HBV DNA sequences in HCC cell lines is possible by the method described, which has potential application for viral genome analysis requiring in situ hybridization.Keywords
This publication has 22 references indexed in Scilit:
- Transformation of NIH/3T3 cells by DNA from a human hepatoma cell line with integrated hepatitis B virus DNAEuropean Journal of Cancer and Clinical Oncology, 1987
- Detection of HBV‐DNA by in situ hybridization using a biotin‐labeled probeJournal of Medical Virology, 1985
- Hepatitis B Virus DNA in Patients with Chronic Liver Disease and Negative Tests for Hepatitis B Surface AntigenNew England Journal of Medicine, 1985
- Hepatitis B Viral Nucleotide Sequences in Non-A, Non-B or Hepatitis B Virus-Related Chronic Liver Disease†Hepatology, 1984
- Establishment of a cell line (HCC‐M) from a human hepatocellular carcinomaInternational Journal of Cancer, 1983
- Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probesVirology, 1983
- Detection of Hepatitis B Virus-specific DNA in the Genomes of Human Hepatocellular Carcinoma and Liver Cirrhosis TissuesJournal of General Virology, 1981
- Detection of hepatitis b virus dna sequences in infected hepatocytes by in situ cytohybridisationJournal of Medical Virology, 1981
- Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell lineNature, 1979
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977