病毒感染相关基因微阵列的制备及其在HBV感染应答基因筛选中的应用

为筛选乙型肝炎(乙肝)病毒(HBV)感染应答基因,探讨HBV感染分子机理,采用生物信息学分析、筛选宿主细胞中与乙肝病毒、丙型肝炎(丙肝)病毒、流行性感冒(流感)病毒等感染密切相关的基因,设计并合成寡核苷酸探针,制备了含231种病毒感染相关基因的寡核苷酸微阵列.利用此微阵列比较HepG2细胞、HepG2.2.15细胞之间的基因表达谱差异,筛选乙肝病毒感染候选应答基因,从分子水平对乙肝病毒感染作用机理进行初步研究.制备的病毒感染相关基因表达谱微阵列的监测结果显示,阳性对照和看家基因探针出现较强信号,空白点样液和阴性对照探针未出信号,大部分基因探针信号强度在可分析范围内,上矩阵和下矩阵反映的基因表达情况一致,证明微阵列的特异性、敏感性、重复性都较好.HepG2.2.15与HepG2细胞基因表达谱比较结果显示,28个宿主基因在HepG2.2.15细胞中高表达,包括ASGR1、AFP、Fibronectin、APOC等基因;4个基因低表达,包括RRM1、ICSBP等基因.初步筛选获得HBV感染候选应答基因.此结果表明,制备的微阵列敏感性、特异性、重复性好,可为研究病毒宿主相互作用关系提供技术平台,应用此微阵列筛选获得的HBV候选应答基因可为揭示HBV感染的分子致病机理提供新的信息,为抗HBV药物研究提供潜在的作用靶点.

Preparation of Oligonucleotide Microarray for Expression Detection of Virus-infection- associated Genes and Its Application in Screening Responsive Genes against HBV Infection

To screen the responsive genes in host with HBV infection and to study the pathology of HBV,the oligonucleotide microarray consisting of 231 kinds of virus-infection-associated genes was constructed.The standards of quality control of it were founded at the same time.With this microarray,the differential gene expression in comparing HepG2.2.15 cell,which is derived from HepG2 cells and produces all HBV proteins, with HepG2 cell was analysed.The mechanism of HBV infection was studied at the molecular level.Our results of quality detecting indicated that the in-house microarray for expression detection of virus-infection-associated genes has high specificity,good sensitivity and stability,etc.Results of microarray for detecting differential expression showed that 28 genes were up-regulated in HepG2.2.15 cells,including ASGR1,AFP,Fibronectin and APOC, 4 genes were down-regulated in HepG2.2.15 cells including RRM1 and ICSBP.These genes may be the responsive genes for HBV infection.In conclusion,our in-house microarray for expression detection of virus-infection-associated genes applys a good technical platform for the study of virus-host interaction.The possible responsive genes to HBV infection derived from this microarray offers new information to the exploitation of pathogenesis of HBV and shows abundant potential targets for the treatment of HBV infection.