人耐药肺腺癌A549/DDP细胞株的差异蛋白质组学  

Differential proteomic analysis of drug resistant A549/DDP cell lines in human lung adenocarcinoma

作　　者：魏瑞[1] 谢云[2] 杨丁懿[1] 贺礼理[1] 彭芳[3] 申良方[1] 

机构地区：[1]中南大学湘雅医院肿瘤科,长沙410008 [2]广东省珠海市人民医院血液肿瘤科,广东珠海519000 [3]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008

出　　处：《中南大学学报（医学版）》2010年第8期854-860,共7页Journal of Central South University ：Medical Science

基　　金：湖南省科技厅科技计划项目(2010SJ3039);湖南省教育厅重点学科建设基金(2009)~~

摘　　要：目的:建立A549与A549/DDP两组细胞株的双向凝胶电泳图谱,识别并鉴定其差异表达的蛋白质,筛选肺腺癌耐药相关蛋白质。方法:收集A549与A549/DDP两组细胞株的总蛋白质,应用二维凝胶电泳(two-dimensional gel electrophoresis,2-DE)进行分离应用,图像分析识别差异表达的蛋白质点,采用基质辅助激光解吸电离飞行时间质谱鉴定差异蛋白质点。使用Western免疫印迹对其中4个蛋白质进行验证。结果:建立了A549与A549/DDP细胞蛋白质的2-DE图谱,识别了40个差异表达的蛋白质点,鉴定了23个差异表达蛋白质。Western免疫印迹证实了差异蛋白质热休克蛋白β1,膜联蛋白A4,丝切蛋白l和波形蛋白在A549与A549/DDP细胞中的差异表达水平。结论:23个差异表达蛋白质为研究肺腺癌耐药机制提供了实验依据。Objective To establish 2-dimensional electrophoresis（2-DE） graph of A549 and A549/DDP cell lines,to identify the differentially expressed proteins,and to screen multidrug resistance（MDR） related proteins in human lung adenocarcinoma.Methods The total proteins of A549 and A549/DDP cells were obtained,and were extracted and separated by 2-DE.PDQuest software was applied to analyze the 2-DE images,and the differential proteins of the 2 types of cells were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF-MS）.Western blot was used to determine the expression levels of the 4 proteins.Results We established 2-DE maps of total proteins from A549 and A549/DDP.A total of 40 differential protein spots in the 2 cell lines were found,and 23 differential expression proteins were identified by MALDI-TOF-MS.Western blot showed that heat shock protein beta-1,annexin A4,cofilin l,vimentin were differential expression proteins in A549 and A549/DDP,which was consistent with the results of the comparative proteomic analysis.Conclusion The 23 differential expression proteins in human lung adenocarcinoma are useful for studying the MDR mechanism of lung adenocarcinoma.

关 键 词：肺腺癌 二维凝胶电泳 蛋白质组学 免疫印迹 

分 类 号：R734.2[医药卫生—肿瘤]