表皮生长因子受体调控的人鼻咽癌细胞系CNE2分泌蛋白质的筛选  

Screening of EGFR-regulated Secreted Proteins in Human NPC Cell Line CNE2

作　　者：梁可[1] 陈主初[1] 易红[1] 李建玲[1] 张鹏飞[1] 李茂玉[1] 李萃[1] 冯雪萍[1] 彭芳[1] 肖志强[1] 

机构地区：[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室

出　　处：《生物化学与生物物理进展》2007年第1期100-106,共7页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展计划(973)(2001CB510207);教育部跨世纪优秀人才培养计划(教育部教技函[2002]48号)基金资助项目.~~

摘　　要：为筛选表皮生长因子受体(epidermalgrowthfactorreceptor,EGFR)调控的鼻咽癌(nasopharyngealcarcinoma,NPC)细胞的分泌蛋白质,揭示EGFR在NPC发病中的作用机制,采用无血清培养法培养NPC细胞系CNE2,并用转化生长因子(transforminggrowthfactor-α,TGF-α)刺激CNE2细胞24h作为实验组,对照组CNE2细胞不用TGF-α刺激.超滤法脱盐并浓缩两组细胞的培养上清制备分泌蛋白,采用双向凝胶电泳技术(two-dimensionalelectrophoresis,2-DE)分离两组细胞的分泌蛋白,PDquest图像分析软件识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(matrix-assistedlaserdesorption/ionizationtimeofflightmassspectrometry,MALDI-TOF-MS)鉴定差异表达蛋白.建立了实验组和对照组CNE2细胞分泌蛋白的2-DE图谱,图像分析识别了22个差异蛋白质点,质谱鉴定了8个非冗余蛋白质,其功能涉及肿瘤细胞侵袭转移、细胞凋亡和增殖,为进一步揭示EGFR在NPC发病中的作用及其机制奠定了基础.In order to screen EGFR-regulated secreted proteins in human nasopharyngeal carcinoma（NPC）, and to reveal the role and mechanism of epidermal growth factor receptor（EGFR） in the pathogenesis of NPC. NPC cell line CNE2 cells were cultured in serum-flee medium and stimulated by transforming growth factor-α （TGF-α） for 24 h in experimental group. Control CNE2 cells were cultured at the same condition but without TGF-α stimulation. The culture medium of control and experimental cells was desalted and concentrated through ultrafiltration to prepared the total secreted proteins. Two-dimensional gel electrophoresis （2-DE） was used to separate the secreted proteins of control and experimental cells, PDQuest software was applied to analyze 2-DE images, and the differential protein spots between the control and experimental cells were identified by desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. The 2-DE patterns of the secreted proteins of TGF-α stimulated and un-stimulated CNE2 cells were established, 22 differential proteins spots between the two groups of cells were found, and 8 non-redundant proteins were identified with MALDI-TOF-MS, the functions of which were involved in invasion, metastasis, apoptosis and proliferation of cancer cells. The data will be valuable for further to study the role and mechanism of EGFR in the pathogenesis of NPC.

关 键 词：鼻咽癌 CNE2细胞 表皮生长因子受体(EGFR) 分泌蛋白质组 二维凝胶电泳 质谱 

分 类 号：Q50[生物学—生物化学] Q51