人肺腺癌细胞株A549和A549/DDP的比较蛋白质组学研究

Comparative proteomic analysis of human lung adenocarcinoma A549 and A549/DDP cells

目的:比较人肺癌细胞株A549和顺铂(cisplatin,DDP)耐药细胞株A549/DDP中的蛋白表达差异。方法:DDP诱导A549细胞建立DDP耐药细胞株A549/DDP。应用蛋白质组学技术分离鉴定A549和A549/DDP细胞中差异表达的蛋白质,通过real-timePCR、蛋白质印迹法和免疫细胞化学法对部分差异蛋白进行验证;应用生物学信息检索分析差异蛋白的功能。结果:A549和A549/DDP细胞中有8个蛋白质点的表达差异>5倍,分别为POTE、FH(fumarate hydratase)、PDE(phosphodiesterase)、AKR1C1(aldo-keto reductase family1,memberC1)、DDH2(dihydrodiol dehydrogenase 2)、S100A10、prefoldin subunit2和核内转运蛋白,这些蛋白与细胞代谢、凋亡、增殖、解毒和信号转导等有关。Real-timePCR、蛋白质印迹法和免疫细胞化学法验证结果与蛋白质组学研究结果一致。结论:鉴定得到的A549和A549/DDP细胞中的差异蛋白为研究肺癌细胞DDP耐药提供新依据。

Objective：To compare the differentially expressed proteins between A549 cells and DDP（cisplatin）-resistant A549/DDP cells.Methods：A549 cells were exposed to DDP for developing the DDP-resistant A549/DDP cells.The differentially expressed proteins in the A549 and A549/DDP cells were separated and identified by proteomics approach.A part of the differentially expressed proteins were validated by real-time PCR,Western blotting and immunocytochemistry.The functions of these proteins were analyzed by bioinformatics method.Results：There were 8 differentially expressed proteins including POTE,FH（fumarate hydratase）,PDE（phosphodiesterase）,AKR1C1（aldo-keto reductase family 1 member C1）,DDH2（dihydrodiol dehydrogenase 2）,S100A10,prefoldin subunit 2,and karyopherin beta2-Ran GppNHp nuclear transport complex between A549 and A549/DDP cells,and the difference in expression was more than 5-fold.These proteins were related with cellular metabolism,apoptosis,cell proliferation,detoxification and signal transduction.The validation results of real-time PCR,Western blotting and immunocytochemistry were consistent with the results of proteomic analysis.Conclusion：The differentially expressed proteins between A549 and A549/DDP cells provide some new clues for the study of DDP-resistant lung cancer cells.