CAJ | 학술논문

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.
목적 분석전렬선상피세포적막단백질구성.방법 장무전렬선병사적시검전렬선표본상규제비빙동조직절편,통과격광포획현미절할(LCM)종조직빙동절편중절취전렬선상피세포,Shotgun-MS기술분석막단백질조학구성.결과 격광포획현미절할가정학유효지분리전렬선상피세포,기동질성＞95%;재엄격적과려삼수조건하(당Charge+1,Xcorr≥1.9;당Charge+2,Xcorr≥2.2;당Charge+3,Xcorr≥3.75;기중DelCN≥0.1),정상전렬선상피세포중감정출1164개단백,기중799개단백경과기인본체평주(GOA)현시위이지세포조분,기여위미지세포조분.재이지세포조분중377(49.15%)개단백위막단백혹자막상관단백.제료이지적여막상관적단백,흔다신적단백야피감정,기중포괄가설단백화일사cDNA서렬.결론 Shotgun-MS결합LCM기술가이유효분석전렬선적막단백질구성;유조우정상전렬선세포막단백질표체고적완선.
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.