CAJ | 학술논문

目的研究人肺腺癌细胞系A549与人正常支气管上皮细胞系16HBE线粒体蛋白质组的差异表达.方法传代培养细胞系A549及16HBE,用线粒体提取试剂盒获取细胞线粒体蛋白质,进行双向凝胶电泳,运用液相色谱串联质谱分析技术筛选出A549和16HBE细胞系线粒体间表达水平显著差异的蛋白,所得结果通过Data Analysis软件标峰,用MASCOT进行结果搜索和数据分析.结果双向电泳结果显示A549、16HBE细胞系线粒体存在差异的蛋白质点共41个,3倍以上差异的16个,A549细胞系中表达上调的15个,表达下调的26个,其中3倍以上差异表达上调的7个,表达下调的9个.运用液相色谱串联质谱技术鉴定出A549细胞系线粒体表达上调的蛋白质2个:AAA+ ATP酶家族结构域蛋白3B、tRNA鸟嘌呤糖基转移酶,表达下调的蛋白质7个:热休克蛋白75、复合物Ⅲ亚基1、复合物Ⅲ亚基2、鸟氨酸氨基转移酶、异柠檬酸脱氢酶亚基α、SLP-2、抗增殖蛋白.结论应用亚细胞蛋白质组学方法,鉴定出肺腺癌细胞系线粒体差异表达蛋白,为阐明肺腺癌发生的分子机制、筛选早期诊断标志物提供了有益的线索.
목적 연구인폐선암세포계A549여인정상지기관상피세포계16HBE선립체단백질조적차이표체.방법 전대배양세포계A549급16HBE,용선립체제취시제합획취세포선립체단백질,진행쌍향응효전영,운용액상색보천련질보분석기술사선출A549화16HBE세포계선립체간표체수평현저차이적단백,소득결과통과Data Analysis연건표봉,용MASCOT진행결과수색화수거분석.결과 쌍향전영결과현시A549、16HBE세포계선립체존재차이적단백질점공41개,3배이상차이적16개,A549세포계중표체상조적15개,표체하조적26개,기중3배이상차이표체상조적7개,표체하조적9개.운용액상색보천련질보기술감정출A549세포계선립체표체상조적단백질2개:AAA+ ATP매가족결구역단백3B、tRNA조표령당기전이매,표체하조적단백질7개:열휴극단백75、복합물Ⅲ아기1、복합물Ⅲ아기2、조안산안기전이매、이저몽산탈경매아기α、SLP-2、항증식단백.결론 응용아세포단백질조학방법,감정출폐선암세포계선립체차이표체단백,위천명폐선암발생적분자궤제、사선조기진단표지물제공료유익적선색.
Objective To investigate the differential mitochondrial protein expressions in human pulmonary adenocarcinoma cell line A549 and normal bronchial epithelial cell line 16HBE.Methods The cell lines A549 and 16HBE were cultured.The cell mitochondria was isolated by mitochondrial fractionation kit.Total proteins of A549 and 16HBE mitochondria were extracted and subjected to twodimensional polyacrylamide gel electrophoresis (2-DE),and then liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to identify the differential proteins.The results was marked peak by data anlasis software,and then the data and analysis results were searched with MASCOT.Results Comparatively analyzing the 2-DE mitochondrial protein patterns of two cell lines,a total of 41 differentially expressed protein spots were found.15 proteins were high-expressed and other 26 proteins were low-expressed in mitochondria of A549.The difference of 16 spots was more than three times,in which seven proteins were high-expressed and nine proteins were low-expressed.Identified by LC-MS/MS,two proteins were high-expressed which were AAA-TOB3 and TGT,and seven proteins were lowexpressed which were HSP75,Complex Ⅲ subunit 1,Complex Ⅲ subunit 2,OAT,ICDHα,SLP-2,and PHB.Conclusions Using the subcellular proteomic method,the differential expression proteins in pulmonary adenocarcinoma cell line are identified.It will provide a uscful clue to elucidate the molecular mechanism of lung cancer and to screen the early diagnostic markers.