家族不良结局乙型肝炎外周血单个核细胞基因表达谱的建立

目的应用寡核苷酸基因芯片技术建立家族不良结局乙型肝炎病毒(HBV)感染者外周血单个核细胞的基因表达谱。方法在一个家族不良结局HBV感染家族中,选取患者5例,患者正常配偶4 例,提取外周血单个核细胞RNA,与涵盖2.2万个ESTs的寡核苷酸表达谱基因芯片U133A 2.0杂交,通过Affymetrix扫描仪和DNT分析软件比较患病组与对照组外周血单个核细胞基因表达谱,获得基因的相对表达比值。结果在2.2万个ESTs中初筛出55个差异表达基因,表达上调14个,表达下调41个,差异基因主要(57%)参与免疫反应、细胞信号转导、细胞周期、代谢、细胞凋亡及炎症基因。结论筛选出的55个基因,是宿主感染HBV的差异表达基因,或宿主对HBV的易感基因,为差异表达基因功能研究建立框架,为宿主HBV易感性研究提供新的靶点。

Genomic analysis of a familial clustering of chronic hepatitis B patients

OBJECTIVE: To investigate the peripheral blood monocyte (PBMC) gene expression profile in a familial clustering of patients with chronic hepatitis B (CHB). METHODS: cRNA prepared from PBMC in a family with 5 CHB patients and 4 normal controls was hybridized to high-density oligouncleotide arrays (HG-U133A 2.0 Human GeneChips, Affymetrix), which interrogate the expression of approximately 22,000 human ESTs. Primary image obtained from scanning was analysed with a DNT software package. Real-time PCR was employed to confirm the gene chip results. RESULTS: 55 genes out of 22,000 ESTs were identified differently. Among the 55 genes 14 showed increased expression and 41 showed decreased expression in the familial clustering CHB patients compared with those in normal controls. Most of the genes (57%) were involved in immunity, inflammation, apoptosis, signaling transduction, and cell cycle. CONCLUSION: These results suggest that the hosts with this broad range of gene expression alterations are susceptible to hepatic B infection.