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

Differential proteomic analysis of Pten^(L/L)MEFs and Pten^(△/△)MEFs cell lines

作　　者：段瑞峰[1] 李刚[1] 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

出　　处：《西安交通大学学报（医学版）》2008年第3期269-273,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30770652)

摘　　要：目的研究PtenL/LMEFs与Pten△/△MEFs细胞的蛋白表达差异。方法运用双向电泳技术比较PtenL/LMEFs与Pten△/△MEFs细胞的蛋白表达差异,6个差异蛋白进行胶内酶切后运用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)进行质谱分析,肽质量指纹图谱检索数据库后5个蛋白得到鉴定。结果同对照PTEN野生型细胞PtenL/LMEFs相比,磷酸甘油酸变位酶1(phosphoglycerate mutase 1,PGAM1)和肽脯氨酰顺反异构酶C(pepti-dyl-prolyl cis-trans isomerase C,cyclophilin C)在Pten△/△MEFs细胞中表达上调而Transgelin 2蛋白在Pten△/△MEFs细胞中表达下调。经鉴定已知蛋白点B和F都是过氧化物氧化还原酶6(Peroxiredoxin-6),蛋白点B在Pten△/△MEFs细胞中表达而在Pten△/△MEFs细胞中检测不到,蛋白点B和F的分子质量相似而等电点不同,可能是由于翻译后修饰。结论同对照PTEN野生型细胞PtenL/LMEFs相比,PTEN缺失的Pten△/△MEFs细胞有明显的差异性蛋白表达谱。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 PtenL/LMEFs and Pten△/△ MEFs cells. Methods Two-dimensional electrophoresis （2-DE） was employed to compare the differential expression proteins between PtenL/LMEFs and Pten△/△ MEFs cells. Six differential expression proteins were digested in gel by enzyme and the mass of generated peptides was measured by matrix assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）. The data obtained from peptide mass fingerprinting （PMF） were searched using the Internet available database and five proteins were identified. Results Compared with that of PtenElL MEFs, expression level of proteins including phosphoglycerate mutase 1 （PGAM1） and peptidyl-prolyl cis-trans isomerase C （PPIC） was up-regulated, whereas expression level of Transgelin 2 was down-regulated in Pten△/△ MEFs cells. B and F proteins were both identified to be peroxiredoxin-6. They had similar molecular weight but different PI which might be caused by post-translation modification. B protein was only expressed in Pten△/△ MEFs cells. Oonclusion The protein profile of Pten△/△ MEFs cells displayed obvious difference compared to that of PtenElL MEFs cells. 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 肽质量指纹图谱 

分 类 号：R730.2[医药卫生—肿瘤]