基于蛋白质谱鉴定的胰腺癌血清标志物同位素标记相对和绝对定量分析  

Isobaric tags for relative and absolute quantitation analysis on serum markers of pancreatic cancer based on mass spectrometry

作　　者：贾凯[1] 耿超[2] 党学义[1] 

机构地区：[1]山西省肿瘤医院普外科,太原030013 [2]中国医学科学院基础医学研究所、北京协和医学院基础学院、国家医学分子生物学重点实验室,北京100730

出　　处：《肿瘤研究与临床》2017年第2期98-103,共6页Cancer Research and Clinic

基　　金：山西省科技攻关项目（100311099.3）

摘　　要：目的 通过蛋白质组学方法筛选胰腺癌患者的血清差异表达蛋白,探讨其临床意义,为生物标记物的发现及验证提供新的依据.方法 采用同位素标记相对和绝对定量（iTRAQ）技术和二维液相色谱-串联质谱（2D-LC-MS/MS）分析方法,对15例胰腺癌患者与10名健康对照的血清进行差异蛋白分析.采用质谱分析并检索Panther、Mascot及Scaffold数据库,对结果进行Ingenuity Pathway Analysis（IPA）分析.结果 从血清样品中共定量442个蛋白.胰腺癌患者与健康对照者血清中存在差异表达的蛋白有76个,并引起相应生物功能改变.胰腺癌患者转化生长因子β1（TGFβ1）、凋亡蛋白酶激活因子1（APAF1）等蛋白表达较健康对照有所下调,而DNA修复蛋白50（RAD50）的表达明显上调.以上三个蛋白与其他蛋白一起形成紧密作用的网络,影响胰腺癌患者的机体代谢.结论 通过蛋白质组学方法筛选出的胰腺癌患者血清差异表达蛋白可能是潜在的胰腺癌标志物,可为临床早期诊断胰腺癌提供分子依据.Objective To determine the expression of serum proteins in pancreatic cancer patients based on mass spectrometry to screen differential proteins and to find potential molecular biomarkers to prediagnosis of pancreatic cancer. Methods The technique of isobaric tags for relative and absolute quantitation （iTRAQ） and two-dimensional liquid chromatography-tandem mass spectrometry （2D-LC-MS/MS） were used to analyze the serum proteins in 15 pancreatic cancer patients and 10 healthy controls. Screening for serum differential proteins was performed via retrieving Panther, Mascot and Scaffold databases. Ingenuity Pathway Analysis was used to detect the correlation between proteins. Results The expression level of 442 proteins was quantified, and 76 proteins were found to be differentially expressed which changed corresponding biological process. The up-regulation of DNA repair protein 50 （RAD50） and down-regulation of transforming growth factor β1 （TGFβ1） and apoptosis protease-activating factor 1 （APAF1） which were correlated with cell growth and apoptosis were found in pancreatic cancer. A closely interacted network was formed between these three differential proteins and other molecules, which could effect metabolism in pancreatic cancer patients. Conclusion Differential expression of serum proteins screened by iTRAQ in pancreatic cancer patients may be the potential markers of pancreatic cancer, which can provide molecular basis for early diagnosis of pancreatic cancer.

关 键 词：胰腺肿瘤 肿瘤标记 生物学 同位素标记相对和绝对定量 蛋白质组学 血清 

分 类 号：R735.9[医药卫生—肿瘤]