Currently, hepatitis B virus (HBV), upon attaching to human hepatocytes, is considered to interact first with heparan sulfate proteoglycan (HSPG) <i>via</i> an antigenic loop of HBV envelope S protein.
In contrast, in a cell line that constitutively expresses both p.R249S and the hepatitis B virus antigen HBx (PLC/PRF/5), silencing of either p.R249S or HBx by RNA interference slowed down proliferation, with no additive effects when both factors were silenced.
In PLC/PRF/5 and Hep3B cells which express hepatitis B-related genes (HBx, HBc and HBc), treatment with PXD101 resulted in apoptosis without a significant effect on viral gene expression.
In this study we investigated the status of the viral enhancer in two human HCC cell lines, Hep3B and PLC/PRF/5 each containing one or more integrated HBV DNA fragments.
These studies show that DFX inhibits the growth of human hepatocellular carcinoma and hepatoblastoma cell lines regardless of the presence (PLC/PRF/5, Hep3B) or absence (Hep G2) of integrated hepatitis B virus DNA.
The human hepatoma cell PLC/PRF/5 contains cloned genomes of hepatitis B virus, one of which was transfected to mouse cells (LTK-) or to human carcinoma cells (HeLa).
Hepatitis B virus (HBV) sequences integrated in the PLC/PRF/5 cell line (Alexander cells), which was derived from a human primary liver carcinoma, were previously extensively studied.
Tissues of human primary hepatocellular carcinoma (PHC) from six patients infected with hepatitis B virus (HBV) were propagated in nude mice, as well as a strain of hepatitis B surface antigen-positive PHC (PLC/PRF/5).
These systems included four human hepatocellular carcinoma cell lines containing HBV DNA (TONG/PHC, HEp 3B2, PLC/PRF/5 and HA22T/VGH) as well as a mouse and a rat cell line both experimentally transfected with HBV DNA.
The human PLC/PRF/5 hepatoma cell line (the Alexander cell) contains at least seven copies of hepatitis B virus (HBV) DNA integrated in its genome; but it selectively expresses the HBV surface antigen (HBsAg) gene and perhaps low levels of the core gene.
The karyotype of the PLC/PRF/5 (Alexander) human hepatoma cell line was identified at passage +/- 110, prior to attempting in situ hybridization studies to determine the chromosomal localization of the hepatitis B virus (HBV) DNA integration sites.
In situ hybridization of an HBV DNA probe to metaphase chromosomes of the PLC/PRF/5 cell line, followed by statistical analysis, identified three integration sites; these were 15q22-q23, 11q22, and 18q12.
We report here the isolation by molecular cloning and the analysis by heteroduplex and restriction enzyme mapping of seven distinct DNA fragments containing hepatitis B virus (HBV) sequences from genomic DNA of the PLC/PRF/5 human liver carcinoma cell line (the Alexander cell).
The rearrangements of chromosome I are most striking in the Hep 3B and PLC/PRF/5 cell lines, which are derived from human hepatocellular carcinomas and contain integrated copies of the hepatitis B viral genome.
Finally, hepatitis B virus DNA, identified in the nude mouse tumor by molecular hybridization techniques, was compared to PLC/PRF/5 cell line hepatitis B virus DNA.
PLC/PRF/5 cell [32P]RNA was found to hybridize with all restriction fragments of HBV DNA adequately tested, indicating that at least most, and possibly all, of the viral DNA in these cells is transcribed.