Locarnini S, Shaw T, Dean J, Colledge D, Thompson A, Li K, Lemon S M, Lau G G K, Beard M R
Victorian Infectious Diseases Reference Laboratory, 10 Wreckyn Street, North Melbourne, Vic 3051, Australia.
J Clin Virol. 2005 Feb;32(2):113-21. doi: 10.1016/j.jcv.2004.10.002.
The expression of the hepatitis Be antigen (HBeAg) is one of several strategies used by hepatitis B virus (HBV) to ensure persistence. The HBeAg may function as a toleragen in utero and has been shown to regulate the host's immune response.
The aim of this study was to examine the effect of the HBV precore and core protein on cellular gene expression in the hepatoma cell line Huh-7.
Huh-7 cells with tight regulated expression of the HBV core or precore protein were produced using the Tet-Off tetracycline gene expression system. Changes in cellular gene expression in response to core/precore expression compared to Huh-7 cells not expressing the proteins were determined using a commercial high-density oligonucleotide array (Affymetrix Hu95A GeneChip) containing probes for 12,626 full-length human genes.
Analysis of differential mRNA gene expression profiles at 7 days post precore and core expression revealed 45 and 5 genes, respectively, with mRNA changes greater than three-fold. The most striking feature was in Huh-7 cells expressing the precore protein in which 43/45 genes were downregulated 3-11-fold. These included genes that encoded products that regulate transcription/DNA binding proteins, cell surface receptors, cell-cycle/nucleic acid biosynthesis and intracellular signalling and trafficking. The only known gene, which was upregulated encoded a cytoskeletal protein. For the core cell line, 4/5 genes were downregulated 3-15-fold upon core induction and included genes that encoded products that affect intermediary metabolism, cell surface receptors and intracellular signalling. The one gene, which was upregulated was a cytokine gene.
The results of this study show that HBV precore protein has a much greater effect on cellular gene expression in comparison to the core protein, suggesting that core and precore proteins may have diverse effects on cellular functions and equally different roles in modulating HBV pathogenesis.
乙肝e抗原（HBeAg）的表达是乙肝病毒（HBV）确保持续感染所采用的多种策略之一。HBeAg在子宫内可能作为一种耐受原，并且已被证明可调节宿主的免疫反应。
本研究旨在检测HBV前核心蛋白和核心蛋白对肝癌细胞系Huh-7中细胞基因表达的影响。
使用Tet-Off四环素基因表达系统构建可严格调控HBV核心蛋白或前核心蛋白表达的Huh-7细胞。与未表达这些蛋白的Huh-7细胞相比，通过使用包含12,626个全长人类基因探针的商业高密度寡核苷酸阵列（Affymetrix Hu95A基因芯片），确定响应核心蛋白/前核心蛋白表达时细胞基因表达的变化。
在前核心蛋白和核心蛋白表达7天后对差异mRNA基因表达谱进行分析，分别发现45个和5个基因的mRNA变化超过三倍。最显著的特征是在表达前核心蛋白的Huh-7细胞中，43/45个基因下调了3-11倍。这些基因包括编码调节转录/DNA结合蛋白、细胞表面受体、细胞周期/核酸生物合成以及细胞内信号传导和运输的产物的基因。唯一上调的已知基因编码一种细胞骨架蛋白。对于核心蛋白细胞系，在核心蛋白诱导后，4/5个基因下调了3-15倍，包括编码影响中间代谢、细胞表面受体和细胞内信号传导的产物的基因。上调的一个基因是细胞因子基因。
本研究结果表明，与核心蛋白相比，HBV前核心蛋白对细胞基因表达的影响要大得多，这表明核心蛋白和前核心蛋白可能对细胞功能有不同的影响，并且在调节HBV发病机制中具有同样不同的作用。
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