论文部分内容阅读
目的:应用同位素标记相对和绝对定量(iTRAQ)技术标记定量蛋白组技术对胃癌患者唾液的全组蛋白进行鉴定和定量分析,初步获得胃癌患者唾液的蛋白组差异表达图谱。方法:胃癌患者与正常人唾液各10例,各组等量混合后利用iTRAQ 8标试剂标记后的样本,采用Nano-LC-Ms/Ms分离、分析肽段,运用Proteinpilot 4.0软件对蛋白进行鉴定和定量分析,并比较这些蛋白的表达差异。结果:对样品进行了2次Nano-LC-Ms/Ms,标记率>95%,错误发现率<1%,鉴定出符合假阳性率<1%,共鉴定了747个蛋白。与正常对照组相比,胃癌组共出现了2倍以上表达差异的蛋白质21个,其中上调的12个,下调的9个。结论:iTRAQ联合Nano LC-MS/MS技术能高通量地筛选胃癌患者唾液中相关的蛋白,为胃癌患者早期诊断和预后提供可能的生物学标志物或治疗靶点,建立一种无创、简便、快捷实用的检测评估手段。
OBJECTIVE: To identify and quantify the salivary proteins in patients with gastric cancer using iTRAQ technique and quantitative proteome technology. The differential expression profiles of saliva in patients with gastric cancer were preliminarily obtained. Methods: Ten patients with gastric cancer and 10 normal controls were enrolled in this study. The samples were labeled with iTRAQ 8 after mixing in equal amounts. The peptides were separated and analyzed by Nano-LC-Ms / Ms. Proteinpilot 4.0 software was used to identify the proteins And quantitative analysis, and compare the differences in the expression of these proteins. Results: Nano-LC-Ms / Ms was performed twice with the labeling rate> 95% and the false discovery rate <1%. The coincidence false positive rate <1% was identified. A total of 747 proteins were identified. Compared with the normal control group, there were 21 proteins with more than 2-fold difference in expression in gastric cancer group, including 12 up-regulated and 9 down-regulated. Conclusion: iTRAQ combined with Nano LC-MS / MS can screen salivary related proteins in gastric cancer patients with high throughput and provide possible biomarkers or therapeutic targets for early diagnosis and prognosis of patients with gastric cancer. A noninvasive and simple , Fast and practical testing and evaluation methods.