【摘要】 本研究以CNE1和CNE2为材料,采用亚细胞蛋白质组研究方法研究不同分化程度鼻咽癌细胞系的差异蛋白质.首先用Percoll密度梯度离心法获得高纯度质膜,通过双向凝胶电泳分离、PDQuest软件分析后找出在肿瘤细胞中表达变化的蛋白质点,再用基质辅助激光解析电离飞行时间串联质谱(MALDI-TOF/TOF-MS)进行鉴定,共鉴定到9个具有2倍或2倍以上差异的蛋白质.这些表达差异的蛋白质参与了细胞分化、代谢及细胞信号传导过程.我们对其中5个蛋白质进行了实时定量PCR分析,对其中4个蛋白质的表达进行了免疫印迹验证.本试验为研究不同分化程度的鼻咽癌提供了一种蛋白质组研究方法,并且找到了galectin-1、annexinⅡ等一些可能与分化相关的蛋白质.这些数据对于研究鼻咽癌的生物学特性具有非常重要的意义.更多还原

【Abstract】 A subcellular proteomic method was applied to investigate the protein expression profiles of nasopharyngeal carcinoma (NPC) cell lines, CNE1 and CNE2, at various differentiation levels. Plasma membrane (PM) proteins were obtained by Percoll density grade centrifugation and subjected to two-dimensional electrophoresis (2DE) followed by PDQuest software analysis. Nine proteins expressed with more than two folds difference were identified by MALDI-TOF-TOF, of which functions involved in cell differentiation, signal transduction, and metabolism. Half of these proteins, such as galectin-1 and annexin Ⅱ, were analyzed with real-time quantitative PCR or Western blotting. We have tested a proteomic method to study differentiated NPCs at different levels and found several proteins that might be related to their biological characteristics.更多还原