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Mutations in the C-terminus of the X protein of hepatitis B virus regulate Wnt-5a expression in hepatoma Huh7 cells: cDNA microarray and proteomic analyses

Liu, X
; 
Wang, L
; 
Zhang, S
; 
Lin, J
; 
Zhang, S
; 
Feitelson, MA
; 
Gao, H
; 
Zhu, M
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Journal Article
Date
2008-06-01
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Subject
Blotting, Western
 Carcinoma, Hepatocellular
 Cell Line, Tumor
 Electrophoresis, Gel, Two-Dimensional
 Gene Expression
 Humans
 Immunohistochemistry
 Liver Neoplasms
 Mutation
 Oligonucleotide Array Sequence Analysis
 Proteomics
 Proto-Oncogene Proteins
 Reverse Transcriptase Polymerase Chain Reaction
 Trans-Activators
 Transfection
 Viral Regulatory and Accessory Proteins
 Wnt Proteins
 Wnt-5a Protein
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http://hdl.handle.net/20.500.12613/5608
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DOI
10.1093/carcin/bgn111
Abstract
Background: The hepatitis B virus x gene (HBx) is a promiscuous transactivator implicated in the development of hepatocellular carcinoma (HCC). The present study was designed to investigate the molecular events regulated by HBx. Methods: Genomic and proteomic expression profiling was performed in Huh7 HCC cells transfected with HBx mutants with a C-terminal deletion. The gene and protein expression of wingless-type murine-mammary-tumour virus (MMTV) integration site family, member 5A (Wnt-5a) was validated by analyses of reverse transcription-polymerase chain reaction (RT-PCR), real-time RT-PCR, western blot and immunohistochemistry. Results: Differentially expressed genes and proteins were found in the transfected Huh7 HCC cells; most of them were involved in transcriptional regulation, although others including oncogenes or tumor suppressor genes, and molecules involved in cell junctions, signal transduction pathways, metabolism or the immune response were also observed. The expression of the Wnt-5a gene was elevated >10-fold in Huh7 cells transfected with the HBx 3′-30 amino acid deletion mutant. However, the expression was downregulated by the transfection with the HBx 3′-40 amino acid deletion mutant. The changes in Wnt-5a expression were also observed in human HCC tissues, compared with corresponding non-cancerous liver tissues. A negative correlation was found between the expression of Wnt-5a and HBx COOH mutations in HCC tissues. Conclusions: HBx mutants may participate in the development and progression of HCC, at least in part through the Wnt-5a pathway. © The Author 2008. Published by Oxford University Press. All rights reserved.
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Oxford University Press (OUP)
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Carcinogenesis
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