人肺腺癌细胞株A549和A549DDP的比较蛋白质组学研究.pdfVIP

170 　　肿瘤2012 年3 月第32 卷第3 期　TUM OR Vol. 32, March 2012 DOI: 10.3781/j .issn.1000-7431.2012 .03.004 Copyright© 2012 by TUM OR Basic Research 基础研究 · 人肺腺癌细胞株A549和A549/DDP的比较蛋白质组学研究 况　鹏，李雪飞，李　冰，王永生，李嘉瑜，周彩存 同济大学附属上海市肺科医院肿瘤科，上海　200433 [摘要]　目的：比较人肺癌细胞株A 549 和顺铂（cisplatin，DDP）耐药细胞株A 549/ 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-time PCR、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：A 549 和A 549/ DDP 细胞中有8 个蛋白质点的表达差异＞5 倍，分别 为POT E、FH （fumarate hydratase）、PDE （phosphodiesterase）、A KR1C1 （aldo-keto reductase family 1，member C1）、DDH2 （dihydrodiol dehydrogenase 2 ）、S100A 10、prefoldin subunit 2 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-time PCR、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。 [关键词]　癌，非小细胞肺；蛋白质组学；药物耐受性；顺铂 [中图分类号]　R734.2 [文献标志码]　A [文章编号]　1000-7431(2012)03-0170-07 Comparative proteomic analysis of human lung adenocarcinoma A549 and A549/DDP cells KUANG Peng, LI Xue-fei, LI Bing, WANG Yong-sheng, LIJia-yu, ZHOU Cai-cun Department of Oncology, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, China [A BSTRACT]　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 incl