人耐药肺腺癌A549/DDP细胞株的差异蛋白质组学

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。

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

ObjectiveTo 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. MethodsThe 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. ResultsWe 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. ConclusionThe 23 differential expression proteins in human lung adenocarcinoma are useful for studying the MDR mechanism of lung adenocarcinoma.