Liu Tian-Hua, Liu Deng-He, Mo Cui-Ju, Sun Lu, Liu Xiao-Xia, Li Wei, Zhang Shu, Liu Yin-Kun, Guo Kun
Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of EducationShanghai, China; Cancer Research Center, Institutes of Biomedical Sciences, Fudan UniversityShanghai, China.
Department of Clinical Laboratory, First Affiliated Hospital of Guangxi Medical University Nanning, Guangxi, China.
Am J Transl Res. 2016 Oct 15;8(10):4250-4264. eCollection 2016.
Hepatic encephalopathy (HE) as a severe neuropsychiatric complication is commonly present in the end stage of Hepatocellular Carcinoma (HCC). However, widely accepted biomarkers for diagnosing early HE are still absent. Here, we screened glycosylation patterns of serum proteins from Chinese HCC patients with or without early HE by lectin microarray. Then, phaseolus vulgaris erythroagglutinin (PHA-E) as a lectin binding with bisecting GlcNAc structure which was significantly decreased in sera from Chinese HCC patients with early HE, was chosen to perform lectin affinity chromatography, following by in-gel digestion, Mass Spectrometry (MS) analysis and bioinformatics analysis. Here we found, 13 lectins showed statistically significant reduction suggesting GalNAc, terminal α-1,3 Man, bisecting GlcNAc, (GlcNAc)n, O-GlcNAc, Neu5Ac, tetra-antennary complex-type N-glycan and GalNAc α/β1-3/6 Gal were decreased in serum glycoproteins from Chinese HCC patients with early HE. Furthermore, a total of 141 PHA-E-associated glycoproteins were identified in MS, of which 12 serum glycoproteins only in Chinese HCC patients without early HE and 26 serum glycoproteins only in Chinese HCC patients with early HE. In addition, bioinformatics analysis revealed the PHA-E-associated serum glycoproteins only in Chinese HCC patients with early HE might be related to early HE occurrence through p38 MAPK signaling pathway and MAPK/ERK signaling pathway. Collectively, this was the first glycomics study of serum proteins in HCC patients with early HE and it could provide a database for discovering and developing serum biomarkers to identify and predict early HE in HCC patients.
肝性脑病（HE）作为一种严重的神经精神并发症，常见于肝细胞癌（HCC）终末期。然而，目前仍缺乏广泛认可的早期HE诊断生物标志物。在此，我们通过凝集素芯片筛选了有或无早期HE的中国HCC患者血清蛋白的糖基化模式。然后，选择与平分型GlcNAc结构结合且在中国早期HE的HCC患者血清中显著减少的菜豆红细胞凝集素（PHA-E）进行凝集素亲和层析，随后进行胶内酶解、质谱（MS）分析和生物信息学分析。我们发现，13种凝集素显示出统计学上的显著减少，表明中国早期HE的HCC患者血清糖蛋白中N-乙酰半乳糖胺、末端α-1,3甘露糖、平分型GlcNAc、（GlcNAc）n、O-连接的N-乙酰葡糖胺、唾液酸、四天线复合型N-聚糖和N-乙酰半乳糖胺α/β1-3/6半乳糖减少。此外，MS共鉴定出141种与PHA-E相关的糖蛋白，其中12种血清糖蛋白仅在中国无早期HE的HCC患者中存在，26种血清糖蛋白仅在中国有早期HE的HCC患者中存在。此外，生物信息学分析显示，仅在中国有早期HE的HCC患者中与PHA-E相关的血清糖蛋白可能通过p38丝裂原活化蛋白激酶信号通路和丝裂原活化蛋白激酶/细胞外信号调节激酶信号通路与早期HE的发生有关。总体而言，这是首次对早期HE的HCC患者血清蛋白进行糖组学研究，可为发现和开发血清生物标志物以识别和预测HCC患者的早期HE提供数据库。
