CAJ | 학술논문

本研究以CNEI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,时其中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、annexin Ⅱ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.
본연구이CNEI화CNE2위재료,채용아세포단백질조연구방법연구불동분화정도비인암세포계적차이단백질.수선용Percoll밀도제도리심법획득고순도질막,통과쌍향응효전영분리、PDQuest연건분석후조출재종류세포중표체변화적단백질점,재용기질보조격광해석전리비행시간천련질보(MALDI-TOF/TOF-MS)진행감정,공감정도9개구유2배혹2배이상차이적단백질.저사표체차이적단백질삼여료세포분화、대사급세포신호전도과정.아문대기중5개단백질진행료실시정량PCR분석,시기중4개단백질적표체진행료면역인적험증.본시험위연구불동분화정도적비인암제공료일충단백질조연구방법,병차조도료galectin-1、annexin Ⅱ등일사가능여분화상관적단백질.저사수거대우연구비인암적생물학특성구유비상중요적의의.
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 twodimensional 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 Westem 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.