肝细胞癌患者血清外泌体蛋白质谱及生物信息学分析

Mass spectrometry and bioinformatics analysis of serum exosome proteins in patients with hepatocellular carcinoma

目的探究肝细胞癌患者与健康人血清外泌体中蛋白质组的差异。方法收集肝细胞癌患者与健康者血清各3例,用试剂盒分离外泌体,提取蛋白。采用液相色谱-质谱联用技术对肝细胞癌患者(试验组)与健康者(对照组)血清外泌体蛋白进行定性分析,并利用DAVID网站进行基因本体论功能分析和KEGG通路富集分析。结果试验组总蛋白数860种,对照组总蛋白数454种,在试验组中3个样本均存在的蛋白有90种,对照组3个样本均存在的蛋白有62种,从这些样本中寻找正常或者肝癌患者特异存在的蛋白,试验组有24种,对照组有3种,并选取铜蓝蛋白作为差异蛋白进行验证。对肝细胞癌患者结果开展信号通路、生物过程以及分子功能等方面的测试分析,其中包括了凝血、炎症反应与天然免疫反应等一系列的过程。结论肝细胞癌患者与健康人血清外泌体蛋白有一定的表达差异,其中铜蓝蛋白差异有可能为肝细胞癌早期诊断提供新的候选分子。

Objective To explore the difference of proteome in serum exosomes between patients with hepatocellular carcinoma and normal healthy people.Methods Collect the serum of 3 patients with hepatocellular carcinoma and 3 healthy people respectively,separate the exosome with the kit,and extract the protein.Qualitative analysis of serum exosome proteins in patients with liver cells(experimental group)and healthy people(control group)by liquid chromatography-mass spectrometry.DAVID website was used to analyze the results of gene ontology function and KEGG pathway enrichment.Results There are 860 kinds of total proteins in the experimental group,454 kinds of total proteins in the control group.There are 90 kinds of proteins in the samples of the experimental group,and 62 kinds of proteins in the samples of the control group.From these samples,we could find the proteins that were specific to normal or liver cancer patients.There were 24 kinds in the experimental group,and 3 kinds in the control group.We selected ceruloplasmin as the differential protein for validation.The analyzed results of patients with hepatocellular carcinoma by molecular function,biotogical process and signal pathway methods,is mainly involving natural immune reaction,inflammatory reaction,blood coagulation and other processes.Conclusion There is a certain difference in the expression of serum exocrine proteins between patients with hepatocellular carcinoma and healthy people,and the difference in ceruloplasmin may provide a new candidate molecule for the early diagnosis of hepatocellular carcinoma.