Viral Hepatitis
Copyright ©2006 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Aug 21, 2006; 12(31): 4986-4995
Published online Aug 21, 2006. doi: 10.3748/wjg.v12.i31.4986
Gene expression analysis of primary normal human hepatocytes infected with human hepatitis B virus
Hyun Mi Ryu, Sung Gyoo Park, Sung Su Yea, Won Hee Jang, Young-Il Yang, Guhung Jung
Hyun Mi Ryu, Guhung Jung, School of Biological Sciences, Seoul National University, Seoul, 151-742, Korea
Sung Gyoo Park, School of Biological Sciences and Institute of Microbiology, Seoul National University, Seoul, 151-742, Korea
Sung Su Yea, Won Hee Jang, The Paik-Inje Memorial Institute for Biomedical Science, Inje University, Pusanjin-gu, Pusan 614-735, Korea
Young-Il Yang, Department of Pathology, College of Medicine, Inje University, Paik Hospital, Pusan 614-735, Korea
Author contributions: All authors contributed equally to the work.
Supported by a grant of the Korea Health 21 R&D Project, Ministry of Health and Welfare, Republic of Korea, No. A050145
Correspondence to: Dr. Guhung Jung, Seoul National Univer-sity, Seoul, 151-742, Korea. drjung@snu.ac.kr
Telephone: +82-2-8807773 Fax: +82-2-8721993
Received: October 30, 2004
Revised: November 11, 2004
Accepted: November 23, 2004
Published online: August 21, 2006
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-κB 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-α, 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 pHBV1.2×, a plasmid encoding all HBV messages. Moreover, these three genes participated in HBV-mediated NF-κB activation.

CONCLUSION: During the initial state of HBV infection, hepatocytes facilitate the activation of NF-κB through up regulation of LT-α, TRAF2, and NIK.

Keywords: cDNA microarray; Primary normal human hepatocytes; LT-α; TRAF2; NIK; NF-κB