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Carcinogenesis Advance Access published online on May 13, 2008
Carcinogenesis, doi:10.1093/carcin/bgn111
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Mutations in the carboxyl terminus of the X protein of Hepatitis B Virus regulate Wnt-5a expression in hepatoma Huh7 cells: cDNA microarray and proteomic analyses

Xiaohong Liu1,4, Li Wang1, Shuhui Zhang1, Jing Lin1, Shunmin Zhang1, Mark A Feitelson2, Hengjun Gao3,* and Minghua Zhu1,*

1 Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai 200433, P. R. China
2 Department of Biology, College of Science and Technology, Temple University, 1900 N. 12th Street, Philadelphia, PA 19122, USA
3 National Engineering Center for Biochip, Shanghai 201203, P. R. China
4 Department of Pathology, Jinan Military General Hospital; Jinan, 250031, China

* To whom correspondence should be addressed: Minghua Zhu, Department of Pathology, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433, China Tel :86-21-25074604, Fax : 86-21-25074604 ; e-mail: (mhzhu2000{at}hotmail.com)

Background/Aims: The hepatitis B 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 COOH-terminal deletion. The gene and protein expression of Wnt-5a (wingless-type MMTV integration site family, member 5A) was validated by analyses of 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 tumour 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 more than 10-fold in Huh7 cells transfected with the HBx3’-30 amino acid deletion mutant. However, the expression was down-regulated by the transfection with the HBx3’-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.

Key Words: Hepatocellular carcinoma • Hepatitis B x gene • COOH-terminal deletion mutation • Wnt-5a • cDNA microarray • proteomics

Received January 7, 2008; revised April 27, 2008; accepted May 3, 2008.


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