汉坦病毒感染细胞整合素受体信号转导相关差异表达基因筛选  

Selection of Differentially Expressed Signal Transduction-Associated Genes of Integrin in Hantavirus Infected Cells

作　　者：徐芳玲[1] 杨占秋[1] 程丽[1] 杨继江[1] 刘汉燕[1] 肖红[1] 文利[1] 丁晓华[1] 陈文凯[1] 

机构地区：[1]武汉大学医学院病毒学研究所教育部病毒学重点实验室,湖北武汉430071

出　　处：《中国病毒学》2004年第4期315-319,共5页Virologica Sinica

基　　金：国家自然基金(39870659);湖北省教育厅(301140166)共同资助

摘　　要：通过基因芯片技术观察汉坦病毒感染细胞后整合素受体信号转导相关基因的差异表达,以探讨汉坦病毒进入细胞的分子机制。用汉滩病毒76-118株感染原代人胚肺成纤维细胞,同时设立正常对照细胞,于感染后第6h、24h和96h同时抽提感染组与正常对照组细胞总RNA,利用荧光标记dUTP逆转录制备cDNA探针,与人类受体及信号转导类表达谱芯片杂交,并对Cy3、Cy5荧光信号做扫描分析。结果显示有37项基因出现差异表达,其中31项基因在感染后6h出现差异表达,13项下调、18项上调;但在感染后24h和96h分别只有5项和4项基因出现表达差异;而且整合素信号转导通路中的重要基因如一些蛋白激酶相关基因在感染6h表达增强,丝化增强子1在感染6h和96h表达下调,微管相关蛋白1A在感染6h表达上调、感染24h和96h表达下调。这些进一步说明汉坦病毒感染细胞与整合素有关。To develop the pathogenesis of Hantavirus infection, the differentially expressed signal trans- duction-associated genes of integrin in Hantavirus infected cells were investigated by using cDNA microarray. The primary human embryo pulmonary fibroblasts (HEPFs) were infected with Hantaan 76-118 virus. Total RNAs were extracted from infected and normal control group 6h, 24h and 96h post infection, respectively. During reverse transcription, the cDNAs were labeled by Cy5-dUTP and Cy3-dUTP. The labeled cDNAs hybridized with the Biostar-ⅡH gene chip. After hybridization, the fluorescence signal was analyzed by scanning. It showed that the expression level of most (113/150) Hantavirus receptor and signal transduction-associated genes was normal, however, 37 genes presented obvious change. Among them, expressions of 31 genes changed at 6h post infection. 13 genes were down-regulated, and 18 up-regulated. However, 5 and 4 genes expressed diversely at 24h and 96h post infection respectively. Furthermore, some integrin signal transduction pathway-associated genes such as the gene of some related protein kinases were up-regulated at 6h post infection. The gene of enhancer of filamentation 1 was down-regulated at 6h and 96h post infection. The gene of microtubule-associated protein 1A was up-regulated at 6h post infection, but down-regulated at 24h and 96h post infection. These indicate that Hantavirus infection is related to integrins.

关 键 词：基因芯片 汉坦病毒 受体 信号转导 原代人胚肺细胞 

分 类 号：R346[医药卫生—基础医学]