摘要目的 探索Shotgun蛋白质组学方法筛选表浅性膀胱移行细胞癌的生物标记组,并分析肿瘤恶性变过程中的生物通路.方法 使用激光捕获显微切割技术获得纯化的表浅性膀胱肿瘤细胞及正常移行上皮细胞,二维液相色谱电喷雾串联质谱鉴定标本中的蛋白质表达,蛋白质组学工具分析蛋白质的等电点、相对分子质量、总体平均疏水性以及跨膜结构.基因本体论工具对鉴定蛋白质进行功能聚类,ArrayTrack软件分析差异表达的生物通路,GenMAPP软件实现生物通路可视化.结果肿瘤细胞及正常移行上皮细胞中分别鉴定蛋白质440、218个,差异表达蛋白质388个.蛋白质鉴定数据以及功能分析数据分别储存于http://www.Proteome-SBTCC.org.cn;http://www.Proteome-NHTE.org.cn,267个(68.8%)差异表达蛋白质具有生物学途径注解,分别归属于丝裂原活化蛋白激酶、聚合粘附、氧化磷酸化、胞外基质受体相互作用等通路.结论 正常移行上皮细胞及肿瘤细胞Shotgun策略蛋白质组学数据库被成功构建,为阐明表浅性膀胱肿瘤的发生发展机制及寻找生物标记组提供了理论依据.

abstractsObjective To study the biomarker panel of superficial bladder transitional cell carcinoma(SBTCC)and analyze the biological pathway in tumorigenesis by Shotgun proteomics strategy.Methods Normal urothelium cells and cancer cells were harvested by laser capture microdissection from clinical specimen and the proteomic expression profile was identified by two-dimensional liquid chromatography tandem mass spectrometry.The isoelectric point,molecular weight,grand average of hydropathicity,transmembrane helices were analyzed by using proteomics tools.Gene ontology was used to comment the identified proteins.The pathway analysis was performed by ArrayTrack software,and visualized by GenMAPP.Results There were 440 and 218 proteins expressed in cancer cells and normal cells respectively,among them 388 proteins were differerntially expressed.All the database about identified proteins was deposited in an accessible form to researchers at http://www.Proteome-SBTCC.org.cn and http://www.Proteome-NHTE.org.cn.There were 267(68.8%)differentially expressed proteins which had GO biological process comments.The biological pathwavs of these proteins included MAPK signaling pathway,focal adhesion,oxidative phosphorylation,ECMreceptor interaction,etc.Conclusion Shotgun strategy proteomies database of normal transitional epithelium and SBTCC is successfully constructed.And the basis for the understanding of cell biology and discovery of biomarker panel for SBTCC iS provided.