【摘要】 为了进一步研究鼻咽癌相关基因NAG7的功能 ,NAG7基因编码框的cDNA片段被亚克隆至 pcDNA3 .1(+ )的表达载体 ,通过脂质体转染入鼻咽癌细胞系HNE1。抽提HNE1细胞和已转染了NAG7基因的HNE1细胞总蛋白质 ,用二维凝胶电泳分离蛋白质 ,获得二维凝胶图谱 ,PDQuest软件包分析后 ,确立表达上调的蛋白质斑点进行质谱分析 ,得到的肽质指纹经蛋白质数据库分析鉴定蛋白质。共分析了 12个蛋白质点 ,其中 3个未有质谱结果 ,已鉴定的 9个蛋白质包括生长捕获特异蛋白、DNA结合蛋白、c myc启动子结合蛋白及胱冬肽酶 (caspase) 6等蛋白质。通过对这些蛋白质性质和功能的讨论 ,发现它们参与了细胞周期调控、转录调节及细胞凋亡等重要事件。因此 ,NAG7很可能是通过介导上述蛋白质的表达上调而发挥作用更多还原

【Abstract】 In search of mechanisms of function of NAG7 gene, a powerful new tool for the unambiguous characterization of gel separated proteins is accomplished by the combination of mass spectrometry and sequence database searching. NAG7 , a novel putative tumor suppressor gene, located on 3p25.3, was introduced into HNE1 cells by liposome transfection. We used two dimensional polyacrylamide gel electrophoresis (2 D PAGE) to identify proteins that were overexpressed in NAG7 transfected cells. After staining and image analysis, spots of interest were isolated and subjected to mass spectrometry (MS). We found nine proteins, which up regulated in NAG7 transfected cells and be identified by MS. These proteins included growth arrest specific protein, DNA binding protein, c myc promoter binding protein and caspase 6 etc., which involved in cell cycling, transcription regulation, and apoptosis. NAG7 may exert the function by up regulating the expression of these proteins.更多还原