前列腺上皮细胞的膜蛋白成分分析

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,Xcorr≥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 cDNA 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.