Wei Rui, Xie Yun, Yang Dingyi, He Lili, Peng Fang, Sheng Liangfang
Department of Oncology, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2010 Aug;35(8):854-60. doi: 10.3969/j.issn.1672-7347.2010.08.013.
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.
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.
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.
The 23 differential expression proteins in human lung adenocarcinoma are useful for studying the MDR mechanism of lung adenocarcinoma.
建立A549和A549/DDP细胞系的二维电泳（2-DE）图谱，鉴定差异表达蛋白，筛选人肺腺癌多药耐药（MDR）相关蛋白。
获取A549和A549/DDP细胞的总蛋白，经2-DE进行提取和分离。应用PDQuest软件分析2-DE图像，采用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）鉴定2种细胞的差异蛋白。采用蛋白质免疫印迹法检测4种蛋白的表达水平。
建立了A549和A549/DDP细胞总蛋白的2-DE图谱。在2种细胞系中总共发现40个差异蛋白点，通过MALDI-TOF-MS鉴定出23个差异表达蛋白。蛋白质免疫印迹法显示，热休克蛋白β-1、膜联蛋白A4、丝切蛋白1、波形蛋白是A549和A549/DDP中的差异表达蛋白，这与比较蛋白质组学分析结果一致。
人肺腺癌中的23个差异表达蛋白有助于研究肺腺癌的MDR机制。
