前列腺上皮细胞的膜蛋白成分分析  

Analysis of human normal prostate cells membrane proteins by shotgun-MS coupled with laser capture microdissection

作　　者：李群[1] 李一雷[1] 李伟[1] 杨新宇[2] 王艺昧[1] 李玉林[1] 

机构地区：[1]吉林大学基础医学院病理生物学教育部重点实验室,长春130021 [2]吉林大学第二临床医院普通外科

出　　处：《中华医学杂志》2008年第3期158-161,共4页National Medical Journal of China

基　　金：教育部重点项目基金（104066）

摘　　要：目的分析前列腺上皮细胞的膜蛋白质构成。方法将无前列腺病史的尸检前列腺标本常规制备冰冻组织切片，通过激光捕获显微切割（LCM）从组织冰冻切片中切取前列腺上皮细胞，Shotgun-MS技术分析膜蛋白质组学构成。结果激光捕获显微切割可正确有效地分离前列腺上皮细胞，其同质性〉95％；在严格的过滤参数条件下（当Charge＋1，Xcorr≥1．9；当Charge＋2，Xcorr≥2．2；当Charge＋3，Xeorr≥3．75；其中DelCN≥0．1），正常前列腺上皮细胞中鉴定出1164个蛋白，其中799个蛋白经过基因本体评注（GOA）显示为已知细胞组分，其余为未知细胞组分。在已知细胞组分中377（49．15％）个蛋白为膜蛋白或者膜相关蛋白。除了已知的与膜相关的蛋白，很多新的蛋白也被鉴定，其中包括假设蛋白和一些cDNA序列。结论Shotgun—MS结合LCM技术可以有效分析前列腺的膜蛋白质构成；有助于正常前列腺细胞膜蛋白质表达库的完善。Objective To analyze of membrane proteins of human normal prostate epithelial cells. Methods Laser capture microdissection （LCM） technique was utilized to obtain the epithelial cells of human normal prostate. Shotgun-MS was used to generate protein profiles in the epithelial cells of human normal prostate. Results LCM technique successfully separated the normal prostate epithelial cells with homogeneity more than 95%. Under a stringent filter condition （ charge ＋ 1, Xcorr≥1.9 ; charge ＋ 2, Xcorr≥2. 2; charge ＋3,Xcorr≥3.75; DelCN≥0. 1） , 1164 proteins were identified in the human normal prostate cells, of which 799 had a gene ontology annotation （GOA） indicating a cellular component, others have no GOA terms. Among the GOA terms, 377 （49.15%） were known membrane proteins or membrane associated proteins. In addition to the proteins known to be associated with the membrane, a significant number of novel proteins had also been identified, including several hypothetical proteins and eDNA sequences. Conclusion Shotgun-MS technique coupled with LCM effectively analyzes the proteins of human normal prostate cells, thus helping perfect the complete protein profiles of human normal prostate cells.

关 键 词：前列腺 上皮细胞 膜蛋白质类 

分 类 号：R686[医药卫生—骨科学]