【作者】 荆涛； 董胜国； 孙立江； 刘勇； 李延江； 杨小坤； 董震；

【Author】 The Department of Urology,the Medical School Hospital of Qingdao University,Qingdao,Shandong 266003 China Jing Tao Dong Shengguo Sun Lijiang Liu Yong Li Yanjiang Yang Xiaokun Dong Zhen

【机构】 青岛大学医学院附属医院泌尿外科；

【摘要】 目的检测膀胱移行细胞癌(transitional cell carcmoma of bladder,TCC)患者尿液中差异表达蛋白质,筛选新的肿瘤标记物,进而探讨其与 TCC 发病机理的相关性,以及蛋白质组学方法在膀胱移行细胞癌早期无创诊断方面的临床应用价值。方法选取青岛大学医学院附属医院泌尿外科 TCC 住院病人尿液标本12例,另外选取4 例 TCC 术后病人、4例健康志愿者以及4例肾肿瘤术前病人的尿液标本作为对照。采用蛋白质芯片技术结合表面增强激光解析/离子化．飞行时间-质谱技术(surface enhanced laser desorption/ ionization-time of flight-mass spectrometry,SELDI-TOF-MS)检测各组尿液标本的差异表达蛋白质,所得数据经计算机处理形成模拟图谱,再通过 Biomark Wizard 软件进行方差分析,最后到 Swiss-Prot 蛋白数据库中鉴定筛选出的肿瘤标记物。结果 TCC 组与对照组尿液标本的蛋白质存在差异表达,IMAC-Cu-3蛋白质芯片共发现6 个蛋白及2个蛋白簇表达水平发生变化。2192 Da,3438 Da,4365 Da,5495Da 及8002Da 差异蛋白质在 TCC 组中表达增高,而4134Da 差异蛋白质及3660Da～3830Da、4310Da～5150Da 差异蛋白簇在对照组中表达增高。联合检测3438Da 及8002Da 可使诊断 TCC 的敏感度达83．3%, 特异度达66．7%。3438Da 蛋白峰已被鉴定为α-defensin 蛋白：搜索 Swiss-Prot 蛋白数据库, 4310Da～5150Da 蛋白簇为 gp40蛋白。结论 SELDI-TOF-MS 蛋白质芯片技术是一种快速、简便易行、用量少和高通量分析方法, 能直接筛选出膀胱移行细胞癌患者尿液中相对特异的潜在肿瘤标记物,为蛋白组学方法在膀胱移行细胞癌早期无创诊断、判断预后及探讨发病机制方面的应用开拓了广阔前景。更多还原

【Abstract】 Objective To detect the different protein expression in the urine of bladder transitional cell carcinoma (TCC),pick out new tumor markers,and explore the correlation between those proteins and the pathogenesis of TCC and the clinical application of proteomics methods for the early noninvasive diagnosis of TCC. Methods:Urine samples from 12 TCC patients and the control group (including 4 postoperaive patients with TCC,4 healthy volunteers and 4 patients with renal tumor) were analyzed using surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS) ProteinChip technology.All experimental data were processed by computer to form the simulating maps and the contents of those different proteins would be statistically analyzed.Then the proteins which had been picked out would be identified in Swiss-Prot protein database. Results:Multiple protein changes were reproducibly detected in the urine samples of TCC group and Control group by IMAC-Cu-3 chip,including six potential novel TCC biomarkers and two protein clusters.Among those proteins,2192Da,3438Da,4365Da,5495Da and 8002Da were up-regulated in TCC group,while 4134Da protein and 3660Da-3830Da,4310Da-5150Da protein clusters were up-regulated in Control group.Combination of the 3438Da and 8002Da biomarkers increased significantly the sensitivity for detecting TCC to 83.3% with a specificity of 66.7%.The 3438Da biomarker had been identified as α-defensin,and the 4310Da-5150Da protein cluster was consistent with gp40 protein in the Swiss-Prot protein database. Conclusion:SELDI-TOF-MS ProteinChip technology is a quick,easy-handling and high-throughput analytic method.This technology can directly screen out relatively specific potential biomarkers from the urine of TCC patients,so it has a better clinical feasibility for early non-invasively diagnosing TCC and will provide some helpful information for predicting the prognosis and studying the pathogenesis of TCC.更多还原