非小细胞肺癌患者尿蛋白质组差异表达分析  

Analysis of Differentially Expressed Proteome in Urine from Non-small Cell Lung Cancer Patients

作　　者：陈振岗[1] 刘金波[1] 林凌[1] 谢辉[1] 张文成[1] 张洪博[1] 王广舜[1] 

机构地区：[1]天津医科大学宝坻临床学院肿瘤外科,天津301800

出　　处：《中国肺癌杂志》2015年第3期138-145,共8页Chinese Journal of Lung Cancer

摘　　要：背景与目的筛查非小细胞肺癌(non-small cell lung cancer,NSCLC)患者尿液中差异表达蛋白,确定可用于NSCLC早期诊断、监测预后和治疗评估的生物标记物。方法分别收集40例已病理证实初诊NSCLC患者、8例肺部良性疾病患者和22例健康志愿者的尿液样本。利用0.9%一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dode-cyl sulfate polyacrylamide gel electrophoresis,1D SDS-PAGE)技术和MS-Thermo-Orbitrap-Velos质谱分析仪对NSCLC组和非肿瘤组尿液中蛋白质进行分离、提取及识别,鉴定出NSCLC患者尿液中的差异表达蛋白。应用SPSS 20.0软件中受试者工作特征曲线(receiver operating characteristic curve,ROC)分别对其敏感性、特异性进行分析,并进行实验验证,从而确定出与NSCLC相关的生物标记物。结果 NSCLC患者组和非肿瘤组尿液差异性表达蛋白质集中表现在90 k Da、60 k Da和20 k Da-30 k Da凝胶条带中。在NSCLC患者尿液蛋白分析中发现了4种与NSCLC相关的差异表达蛋白,包括上调蛋白LRG1、CA1和下调蛋白VPS4B、YWHAZ。这4种差异表达蛋白作为独立的NSCLC生物标记物其敏感性较低:LRG1蛋白敏感性83.0%(25/30)、特异性90.0%(18/20);CA1蛋白敏感性60.0%(18/30)、特异性90.0%(18/20);VPS4B蛋白敏感性73.3%(22/30)、特异性90.0%(18/20);YWHAZ蛋白敏感性60.0%(18/30)、特异性95.0%(19/20)。而采用蛋白质组合模式对NSCLC进行筛查、诊断,则其敏感性和特异性分别可高达96.7%(29/30)和85%(17/20)。结论 LRG1、CA1蛋白在NSCLC患者尿液中高表达,而VPS4B、YWHAZ蛋白呈低表达,差异表达蛋白均提示有可能成为用于NSCLC早期筛查、监测预后和治疗评估的生物标记物。LRG1、CA1、VPS4B和YWHAZ尿液蛋白作为单一生物标记物应用于NSCLC筛查和诊断的敏感性较低,而采用蛋白质组合模式明显优于独立模式对NSCLC的筛查和诊断,故蛋白质组合模式在临床诊疗中将更具有良好应用价值和前景。Background and objective Screen differentially expressed proteins in patients with non-small cell lung cancer（NSCLC）, and aim to identify biomarkers for early screening, monitoring prognosis and evaluating therapy of NSCLC. Methods Urinary samples were collected from 40 newly diagnosed NSCLC patients, 8 patients with lung benign disorders and 22 healthy people. 0.9% sodium dodecylsulfate- polyacrylamide gel electrophoresis（1D SDS-PAGE） and MS-Thermo-Orbitrap-Velos were applied to separate, extract and identify proteins in urinary samples from non-neoplastic groups and NSCLC patients, in order to find out differentially expressed proteins in patients with NSCLC. Then, sensitivity and specificity of candidate proteins were tested by certain experiments. Finally, biomarkers related to NSCLC could be determined. Results The differences of urinary proteins between non-neoplastic groups and NSCLC patients mainly focused on 90 k Da, 60 k Da and 20 k Da-30 k Da stripes. Four differently expressed proteins were found in urinary proteins in NSCLC group, including LRG1, CA1（up-regulating proteins） and VPS4 B, YWHAZ（down-regulating proteins）. The sensitivity of these four proteins for biomarker of NSCLC was relatively low when they were used to screen or diagnose independently. The sensitivity and specificity of LRG1 was 83.0%（25/30） and 90.0%（18/20）, respectively; 60.0%（18/30） and 90.0%（18/20） for CA1; 73.3%（22/30） and 90.0%（18/20） for VPS4B; 60.0%（18/30） and 95.0%（19/20） for YWHAZ. However, the sensitivity and specificity would increase to 96.7%（29/30） and 85%（17/20） after the four biomarkers were combined. Conclusion LRG1 and CA1 are abundant in urine in patients with NSCLC, while VPS4 B and YWHAZ are low-abundance proteins. They could be regarded as biomarkers for early screening, monitoring prognosis and evaluating therapy of patients with NSCLC because of differential expression.The sensitivity of the four biomarkers of NSCLC is relatively low when they are used t

关 键 词：肺肿瘤 蛋白质组学 尿液 生物标记物 

分 类 号：R734.2[医药卫生—肿瘤]