Gene expression analysis of primary normal human hepatocytes infected with human hepatitis B virus.
OBJECTIVE
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.


CONCLUSIONS
During the initial state of HBV infection, hepatocytes facilitate the activation of NF-kappaB through up regulation of LT-alpha, TRAF2, and NIK.
Ryu
Hyun Mi
HM
School of Biological Sciences, Seoul National University, Seoul, 151-742, Korea.
Park
Sung Gyoo
SG
Yea
Sung Su
SS
Jang
Won Hee
WH
Yang
Young-Il
YI
Jung
Guhung
G
D018076:DNA, Complementary / Q000378:metabolism; D005786:Gene Expression Regulation; D015972:Gene Expression Regulation, Neoplastic; D006509:Hepatitis B / Q000378:metabolism / Q000473:pathology*; D006515:Hepatitis B virus / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism* / Q000821:virology*; D006801:Humans; D015151:Immunoblotting; D016328:NF-kappa B / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D047992:TNF Receptor-Associated Factor 2 / Q000378:metabolism; D014162:Transfection; D000097782:NF-kappaB-Inducing Kinase
D018076:DNA, Complementary; D016328:NF-kappa B; D017346:Protein Serine-Threonine Kinases; D047992:TNF Receptor-Associated Factor 2; D000097782:NF-kappaB-Inducing Kinase