HCV全基因组培养细胞的比较蛋白组学研究

Comparative Proteomics Analysis of Huh7 Cells Harboring Intact HCV

利用比较蛋白质组技术研究了转染丙型肝炎病毒(Hepatitis C virus,HCV)全基因组的人肝癌细胞系Huh7细胞模型中蛋白质表达谱的变化,建立了Huh7-HCV的双向凝胶电泳蛋白质表达图谱和数据库.通过双向凝胶电泳分离和图像分析,对表达差异2倍以上蛋白质点进行了胶内酶解和MALDI-TOF MS鉴定.得到包括与细胞骨架蛋白、细胞周期、凋亡和信号转导等相关的14个蛋白质,并且用Western blot验证了热休克蛋白70的蛋白质组研究结果.利用HCV全基因组培养系统,采用蛋白质组学技术,为研究HCV病毒和宿主细胞相互作用提供了新的实验数据,为深入研究HCV病毒复制和分子致病机理奠定了基础.

To analyze the protein expression changes of a human liver cell lines Huh7 in the absence and presence of HCV, proteomic techniques were used to establish the protein expression profiles. The differentially expressed proteomes between Huh7 cells and Huh7 cells with HCV were profiled and the 14 differentially expressed proteins were identified via two dimensional electrophoresis and MALDI-TOF-MS. Many proteins, included regucalcin, centriolin and several keratins, were up regulated expression after HCV transfection. Among down-regulated expression proteins, heat shock proteins were very attractive. Subsequently, we randomly identified the proteomics results by the detection of Hsp70 expression levels between Huh7 cells and Huh7 cells with HCV by western blot methods. Our study provides the global information of proteomic alteration of Huh7 cells in the presence of intact HCV and the clues for further understanding of the mechanism of interaction between HCV and host cell.