基于iTRAQ技术的肝癌肝郁证唾液蛋白质组学

Salivary Proteomics of Liver Cancer with Liver-Depression Syndrome Using iTRAQ Techniques

通过同位素标记相对和绝对定量(iTRAQ)技术分析原发性肝癌不同中医证型的差异蛋白表达谱,拟从唾液蛋白质水平阐述原发性肝癌肝郁证的证候本质。共收集肝郁证肝癌患者、非肝郁证肝癌患者以及健康对照组唾液样本各11例。在3组人群共鉴定1296个蛋白;与健康对照组比较,肝郁证组和非肝郁证组分别有390和231个显著差异表达蛋白(差异倍数>1.5或<0.67,p值<0.05)。进一步比较肝郁证组和非肝郁证组的差异蛋白,发现260个蛋白在肝郁证组中特异性表达,包括124个上调蛋白和136个下调蛋白。GO功能分类和KEGG通路分析表明,肝郁证组中特异表达蛋白主要涉及蛋白酶体通路、溶酶体通路、黏附连接通路等。建立了肝郁证肝癌特异的蛋白数据库,筛选出与肝郁证肝癌发生发展可能相关的多个差异蛋白。

The aim of this study is to explore the pathogenesis of hepatocellular carcinoma (HCC) with liver-depression syndrome from the level of salivary protein by analyzing the protein expression profiles of different TCM syndromes of primary HCC with isobar labeled relative and absolute quantitation (iTRAQ) techniques. Eleven saliva samples from HCC patients with liver-depression syndrome,HCC patients without liver-depression syndrome,and healthy controls were collected,respectively. A total of 1 296 proteins were identified among all three groups. Compared with the healthy control group(fold change >1.5 or <0.67 and p value <0.05),390 differentially expressed proteins were detected in HCC patients with liver-depression syndrome and 231 differentially expressed proteins were detected in HCC patients without liver-depression syndrome. Further comparison between the two groups reveals that 260 proteins are specifically expressed in HCC patients with liver-depression syndrome,including 124 up-regulated and 136 down-regulated proteins. Gene Ontology functional classification and Kyoto Encyclopedia of Genes and Genomes pathway analysis show that these proteins are mainly involved in proteasome pathways,lysosomal pathwaysand adhesion pathways. We established the differentially expressed protein database for HCC with liver-depression syndrome. The multiple differentially expressed proteins that might be related to the pathogenesis of HCC were identified.