Liu Xiaohong, Wang Li, Zhang Shuhui, Lin Jing, Zhang Shunmin, Feitelson Mark A, Gao Hengjun, Zhu Minghua
Department of Pathology, Changhai Hospital, Second Military Medical University, 174 Changhai Road, Shanghai 200433, People's Republic of China.
Carcinogenesis. 2008 Jun;29(6):1207-14. doi: 10.1093/carcin/bgn111. Epub 2008 May 13.
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.
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.
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 HBx3'-30 amino acid deletion mutant. However, the expression was downregulated 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.
HBx mutants may participate in the development and progression of HCC, at least in part through the Wnt-5a pathway.
乙型肝炎病毒X基因（HBx）是一种多效性反式激活因子，与肝细胞癌（HCC）的发生发展有关。本研究旨在探究受HBx调控的分子事件。
对转染了C末端缺失的HBx突变体的Huh7肝癌细胞进行基因组和蛋白质组表达谱分析。通过逆转录-聚合酶链反应（RT-PCR）、实时RT-PCR、蛋白质印迹和免疫组织化学分析，验证无翅型鼠乳腺肿瘤病毒（MMTV）整合位点家族成员5A（Wnt-5a）的基因和蛋白表达。
在转染的Huh7肝癌细胞中发现了差异表达的基因和蛋白质；其中大多数参与转录调控，不过也观察到了其他一些基因，包括癌基因或肿瘤抑制基因，以及参与细胞连接、信号转导通路、代谢或免疫反应的分子。在转染了HBx 3'-30氨基酸缺失突变体的Huh7细胞中，Wnt-5a基因的表达升高了10倍以上。然而，转染HBx 3'-40氨基酸缺失突变体后，该表达下调。与相应的癌旁肝组织相比，在人类肝癌组织中也观察到了Wnt-5a表达的变化。在肝癌组织中，Wnt-5a的表达与HBx羧基末端突变之间呈负相关。
HBx突变体可能至少部分通过Wnt-5a途径参与HCC的发生和发展。
