基于鸟枪法蛋白质组学和生物信息学技术对肺鳞癌表达蛋白质谱的分析  

Analysis of expression protein profiles of lung squamous cell carcinoma based on shot gun proteomics and bioinformatics technique

作　　者：南岩东[1] 杨拴盈[2] 田应选[3] 霍淑芬[2] 杜洁[2] 金发光[1] 

机构地区：[1]第四军医大学唐都医院呼吸内科,陕西西安710038 [2]西安交通大学医学院第二附属医院呼吸内科,陕西西安710004 [3]陕西省人民医院老年呼吸科,陕西西安710068

出　　处：《西安交通大学学报（医学版）》2011年第1期10-16,27,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的应用高通量蛋白质组学及生物信息学技术分析肺鳞癌表达蛋白质谱,筛选肺癌标志蛋白质,为肺鳞癌发病机制、早期诊断及治疗提供有价值的线索。方法 6例手术后新鲜肺鳞癌组织,通过激光捕获显微切割(laser capture microdissection,LCM)分离纯化肿瘤细胞并提取可溶性总蛋白;多维液相色谱-离子阱串联质谱技术(也称鸟枪法蛋白质组学,shot gun proteomics)对蛋白质进行分离、鉴定;进一步通过生物信息学工具预测肺鳞癌细胞表达蛋白质的理化性质、分子功能、生物通路及蛋白质相互作用,并筛选潜在的标志蛋白。结果 LCM共收集了60个帽(cap)的感兴趣细胞,平均每个LCM cap捕获感兴趣细胞数约12000个,其同质性大于95%。可溶性总蛋白经多维液相色谱-离子阱串联质谱共鉴定出860个非冗余蛋白质。蛋白质的理化性质包括分子量、等电点、平均疏水性、跨膜结构域、亚细胞定位、转录后修饰和组织分布等经过在线生物信息学工具分析,并通过统计图直观显示;蛋白质生物学功能基于Gene Ontologyc(GO)软件和Kyoto encyclopedia of genes and genomes(KEGG)生物学通路分析,根据其注解,筛选出3个有价值的蛋白质,即分裂素活化蛋白激酶(mitogen-activated protein kinase,MAPK)、粘联素A1(annexin A1,ANXA1)、高移动组蛋白B1(high mobility group protein B1,HMGPB1)可以作为肺鳞癌候选的标志蛋白。结论鸟枪法蛋白质组学可大规模分离、鉴定肿瘤细胞表达蛋白质,基于生物信息学筛选的标志蛋白质可能成为肺癌诊断和治疗的分子靶点。Objective To analyze the expression protein profiles of lung squamous cell carcinoma（SQCC） with shot gun proteomics and bioinformatics technique and to screen SQCC biomarkers in order to provide valuable clues for pathogenesis,early diagnosis and treatment of SQCC.Methods Tumor cells were isolated and purified with laser capture microdissection（LCM） from 6 cases of fresh tissues of lung SQCC after operatoin and the total solution proteins were extracted.Proteins were identified by multiple-dimension liquid chromatography（MDLC） and ion trap tandem mass spectrum technology（shot gun proteomics）.Biomarkers were screened based on prediction of the physio-chemical properties,molecular function,biological pathway and interaction of the expressoin proteins using bioinformatics technique.Results A total of 60 LCM caps were collected and about 12000 cells were harvested from every LCM cap.Altogether 860 non-redundant proteins were identified using shot gun proteomics technique.The physio-chemical properties,including molecular weight（MW）,point of isoelectricity（PI）,grand average of hydropathy（GRAVY）,trans-membrane helices（TMH）,subcellular location,post-transcription modification（PTM） and tissue distribution were analyzed using online bioinformatics tools and werefurther demonstrated through statistical graph.Biological function was analyzed by using gene ontology（GO） software and kyoto encyclopedia of genes and genomes（KEGG） biological pathway.Three valuable proteins,namely,mitogen-activated protein kinase（MAPK）,annexin A1（ANXA1） and high mobility group protein B1（HMGPB1） were screened as candidate biomarkers for lung SQCC according to functoin annotatoin.Conclusion Shot gun proteomics can globally isolate and identify the expression protein of tumor cell.The screened biomarker proteins based on bioinformatics technique may become a molecular target point of diagnosis and treatment of lung cancer.

关 键 词：肺鳞癌 鸟枪法蛋白质组学 生物信息学 标志蛋白质 分裂素活化蛋白激酶 粘联素A1 高移动组蛋白B1 

分 类 号：R734.2[医药卫生—肿瘤]