Pten^(+/+)MEFs与Pten^(-/-)MEFs细胞系的差异表达蛋白质组分析  

Differential Proteomic Analysis of Pten^(+/+)MEFs and Pten^(-/-)MEFs Cell Lines

作　　者：段瑞峰[1] 傅春玲[2] 李刚[1] 杨柳[2] 胡迎春[1] 吴德昌[1] 霍艳英[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,北京100850 [2]重庆医科大学病理学教研室,重庆400016

出　　处：《生物技术通讯》2008年第3期368-371,共4页Letters in Biotechnology

基　　金：国家自然科学基金项目(30770652)

摘　　要：目的:研究PTEN缺失细胞的蛋白表达规律。方法:用双向电泳技术比较Pten+/+MEFs与Pten-/-MEFs细胞的蛋白表达差异;用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)对差异蛋白进行质谱分析;用肽质量指纹图谱检索数据库对差异蛋白进行鉴定;用Northern印迹和Western印迹验证蛋白的差异表达。结果:与Pten+/+MEFs细胞相比,Pten-/-MEFs细胞有明显的差异性蛋白表达谱,Pten-/-MEFs细胞中表达下降的蛋白有铜锌超氧化物歧化酶、过氧化物氧化还原酶5和6等,表达增加的蛋白有低分子量蛋白酪氨酸磷酸酶和丝切蛋白(coffilin)1。结论:PTEN的缺失引起细胞内多种蛋白表达改变,这些表达改变的蛋白可能与PTEN缺失后细胞癌变相关。Objective： To study protein variation between Pten^＋/＋MEFs and Pten^-/-MEFs cells. Methods： Two dimensional electrophoresis was employed to compare the differential expression proteins between Pten^＋/＋MEFs and Pten^-/-MEFs cells. The differential expression proteins were digested in gel by enzyme and the mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry. The data obtained from peptide mass fingerprinting was searched using the internet available database. The expressions of some identified proteins were confirmed using Northern blot and Western blot. Results： The Cu/Zn-SOD, Prx5 and Prx6 were down-regulated while the cofilin 1 and low molecular weight protein tyrosine phosphatase were up-regulated in Pten^-/-MEFs cells. The expression of Cu/ZnSOD, Prx5 and Prx6 was confirmed using Northern blot and Western blot. Conclusion： The protein profile of Pten^-/-MEFs cell lines displayed obviously difference compared to that of Pten^＋/＋MEFs cell lines. The results implied that various distinct different proteins might lead to cancer.

关 键 词：PTEN 双向电泳 基质辅助激光解吸/电离飞行时间质谱 

分 类 号：Q26[生物学—细胞生物学] Q503