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ABSTRACT: Aim
To find the relationship between hepatitis B virus (HBV) and hepatocytes during the initial state of infection by cDNA microarray.Methods
Primary normal human hepatocytes (PNHHs) were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA (cDNA) with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were produced and some of them were confirmed by RT-PCR, immunoblot analysis, and NF-kappaB luciferase reporter assay.Results
From the cDNA microarray, we obtained 98 differentially regulated genes. Of the 98 genes, 53 were up regulated and 45 down regulated. Interestingly, in the up regulated genes, we found the TNF signaling pathway-related genes: LT-alpha, TRAF2, and NIK. By using RT-PCR, we confirmed the up-regulation of these genes in HepG2, Huh7, and Chang liver cells, which were transfected with pHBV1.2x, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV-mediated NF-kappaB activation.Conclusion
During the initial state of HBV infection, hepatocytes facilitate the activation of NF-kappaB through up regulation of LT-alpha, TRAF2, and NIK.
SUBMITTER: Ryu HM
PROVIDER: S-EPMC4087401 | biostudies-literature | 2006 Aug
REPOSITORIES: biostudies-literature
Ryu Hyun Mi HM Park Sung Gyoo SG Yea Sung Su SS Jang Won Hee WH Yang Young-Il YI Jung Guhung G
World journal of gastroenterology 20060801 31
<h4>Aim</h4>To find the relationship between hepatitis B virus (HBV) and hepatocytes during the initial state of infection by cDNA microarray.<h4>Methods</h4>Primary normal human hepatocytes (PNHHs) were isolated and infected with HBV. From the PNHHs, RNA was isolated and inverted into complement DNA (cDNA) with Cy3- or Cy5- labeled dUTP for microarray analysis. The labeled cDNA was hybridized with microarray chip, including 4224 cDNAs. From the image of the microarray, expression profiles were ...[more]