MDCK细胞感染犬细小病毒后的差异表达谱分析

通过了解MDCK细胞感染犬细小病毒(CPV)后基因表达水平的差异,研究病毒对细胞的致病作用以及细胞抵御病毒感染的机制。利用表达谱基因芯片技术,分析持续感染犬细小病毒的MDCK细胞基因表达水平的变化情况,并用Real-Time PCR技术加以验证。结果显示,获得了359个差异大于1.5倍的表达基因(P0.05),占总基因数的1.53%,其中193个上调表达(0.84%),166个下调表达(0.69%)。对差异基因进行GO功能聚类分析,小部分涉及免疫应答、生长周期调控、信号转导和蛋白酶活性,其他大部分基因功能未知。利用Real-Time PCR随机验证5个基因在持续感染CPV后的差异表达,其结果和芯片杂交的结果一致。表明建立了MDCK细胞持续感染CPV后的差异表达谱,初步了解到CPV对宿主细胞的制约作用,引起部分细胞增殖调控相关基因发生了表达下调,以及细胞对感染的积极应答反应,部分免疫反应基因、肽链内切酶活性基因等发生了上调表达,从而为探索病毒的致病机理和宿主的抗病毒途径提供了试验基础。

Genomic expression profiling of Madin-Darby canine kidney(MDCK) cells with persistent canine parvovirus infection

In order to understand the differences in gene expression of MDCK cells infected with canine parvovirus(CPV),and elucidate the mechanism of virus infection and cellular antivirals,microarray technologies were used to analyze the change of genomic expression in MDCK cells with CPV persistent infection,and the canine genomic expression profiling was established.Real-Time PCR technology was used to further verify the results of microarray.359 genes with 1.5-fold difference of expression(1.53% of total number of genes) were obtained,in which 193 genes were up-regulated(0.84%),and 166 down-regulated(0.69%).GO functional cluster analysis showed that some genome altered involved in immune response,the growth cycle regulation or signal transduction and protease activity,but some other functions were unknown.Five randomly altered expression genes verified by real-time PCR showed the results were consis-tent with the microarray analysis.A genomic expression profiling of MDCK cells with persistent infection CPV was established,and the restriction role of CPV to host cells and the positive responses of host cells to CPV were initially understood,which showed that parts of the proliferation-regulated genes were expressed down,and some of the immune response genes,endopeptidase activity genes were up-regulated.The results of this study provided a basis of further research for understanding the molecular pathogenesis of CPV infection and the host antiviral mechanism.