鼻咽癌转移相关的分泌蛋白质的筛选  

Identification of metastasis-related secretory proteins in human nasopharyngeal carcinoma cells

作　　者：向亚莉[1] 易红[1] 李茂玉[1] 张鹏飞[1] 李萃[1] 彭芳[1] 阮林[1] 陈主初[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：《国际病理科学与临床杂志》2007年第5期369-375,共7页Journal of International Pathology and Clinical Medicine

基　　金：国家重点基础研究发展计划(973)资助项目(2001CB510207);湖南省科技厅重点科研项目(06SK2004)~~

摘　　要：目的:筛选鼻咽癌(nasopharyngeal carcinoma,NPC)转移相关的分泌蛋白质,为阐明NPC转移机制以及筛选NPC转移分子标志物提供实验依据。方法:应用二维凝胶电泳(two-dimensional electrophoresis,2-DE)技术分离一对来自同一亲本,具有不同转移潜能的NPC细胞系5-8F和6-10B细胞的分泌蛋白质,图像分析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(MALDI-TOF-MS)和电喷雾-四极杆-串联质谱(ESI-Q-TOFMS/MS)对差异表达的蛋白质点进行鉴定。Western印迹法检测差异分泌蛋白质nm23-H1在两株细胞中的表达水平。结果:建立了5-8F和6-10B分泌蛋白质的2-DE图谱,质谱分析鉴定出14个非冗余的分泌蛋白质,其中Oncop-rotein18等蛋白质在高转移潜能NPC细胞系5-8F中的表达水平高于无转移潜能的NPC细胞系,而nm23-H1等蛋白质在5-8F细胞系中的表达水平低于6-10B细胞系。Western印迹分析证实了nm23-H1在5-8F和6-10B细胞分泌蛋白质中的差异表达水平。结论:所鉴定的14个非冗余的差异分泌蛋白质为研究NPC转移机制以及筛选NPC转移分子标志物提供了实验依据。Objective To screen metastasis-related secretory proteins in nasopharyngeal carcinoma（NPC）,and provide experimental evidences for elucidating the metastatic mechanism of NPC and screening the metastatic biomarkers of NPC.Methods The secretory proteins of a pair of NPC cell lines 5-8F and 6-10B with different metastatic potential and the same genetic background were separated by two-dimensional gel electrophoresis（2-DE）,respectively.PDQuest software was applied to analyze 2-DE images,the differentially expressed protein spots between 5-8F and 6-10B were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS） and electrospray ionization-quadrupole time-of-flight MS/MS（ESI-Q-TOF MS/MS）.The expression levels of differential protein nm23-H1 in the secretory proteins of two cell lines were detected by Western blot.Results 2-DE maps of secretory proteins from 5-8F and 6-10B were established.Fourteen non-redundant differential expression proteins were identified by mass spectrometry.Some differential proteins such as oncoprotein 18 were significantly higher in highly metastatic 5-8F than those in non-metastatic 6-10B,and other differential proteins such as nm23-H1 were significantly lower in highly metastatic 5-8F than that in non-metastatic 6-10B.The differential expression levels of nm23-H1 in secretory proteins of 5-8F and 6-10B were confirmed by Western blot.Conclusion Fourteen non-redundant differential expression proteins will be valuable for studying the metastatic mechanism of NPC and screening the metastatic biomarkers of NPC.

关 键 词：鼻咽癌 转移 分泌蛋白质 双向凝胶电泳 质谱 

分 类 号：R739.63[医药卫生—肿瘤]