Alteration of N-glycan expression profile and glycan pattern of glycoproteins in human hepatoma cells after HCV infection

• Published in: Biochimica et biophysica acta. General subjects Vol. 1861; no. 5; pp. 1036 - 1045
• Main Authors: Xiang, Tian, Yang, Ganglong, Liu, Xiaoyu, Zhou, Yidan, Fu, Zhongxiao, Lu, Fangfang, Gu, Jianguo, Taniguchi, Naoyuki, Tan, Zengqi, Chen, Xi, Xie, Yan, Guan, Feng, Zhang, Xiao-Lian
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2017
• Subjects: agglutinins; Annexin A2 - metabolism; biomarkers; Carcinoma, Hepatocellular - metabolism; Carcinoma, Hepatocellular - virology; Cell Line; Fucose - metabolism; Fucosyltransferases - metabolism; gene expression regulation; glycoproteins; Glycoproteins - metabolism; glycosylation; Glycosyltransferase expression analysis; glycosyltransferases; Glycosyltransferases - metabolism; heat shock proteins; Hepacivirus - pathogenicity; hepatitis C; Hepatitis C - metabolism; Hepatitis C - virology; Hepatitis C virus; Hepatitis C virus (HCV); hepatocytes; hepatoma; Humans; Lectin microarray; lectins; Lectins - metabolism; Lens culinaris; life cycle assessment; Liver - metabolism; Liver - virology; liver cirrhosis; Liver Cirrhosis - metabolism; Liver Cirrhosis - virology; Liver Neoplasms - metabolism; Liver Neoplasms - virology; Mass spectrometry; Membrane Glycoproteins - metabolism; messenger RNA; microarray technology; N-glycan; Plant Lectins - metabolism; Polysaccharides - metabolism; quantitative polymerase chain reaction; reverse transcriptase polymerase chain reaction; Western blotting; α1,6-fucosyltransferase 8 (FUT8); ECA; Lectin microarray; α1,6-fucosyltransferase 8 (FUT8); ANXA2; Glycosyltransferase expression analysis; HCC; AFP; LCA; Hepatitis C virus (HCV); HCVcc; HSP90B1; NS3; FUT8; PFN-1; POFUT1; HCV; N-glycan; Mass spectrometry; ACA
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.02.014

Hepatitis C virus (HCV) infection causes chronic liver diseases, liver fibrosis and even hepatocellular carcinoma (HCC). However little is known about any information of N-glycan pattern in human liver cell after HCV infection. The altered profiles of N-glycans in HCV-infected Huh7.5.1 cell were analyzed by using mass spectrometry. Then, lectin microarray, lectin pull-down assay, reverse transcription-quantitative real time PCR (RT-qPCR) and western-blotting were used to identify the altered N-glycosylated proteins and glycosyltransferases. Compared to uninfected cells, significantly elevated levels of fucosylated, sialylated and complex N-glycans were found in HCV infected cells. Furthermore, Lens culinaris agglutinin (LCA)-binding glycoconjugates were increased most. Then, the LCA-agarose was used to precipitate the specific glycosylated proteins and identify that fucosylated modified annexin A2 (ANXA2) and heat shock protein 90 beta family member 1 (HSP90B1) was greatly increased in HCV-infected cells. However, the total ANXA2 and HSP90B1 protein levels remained unchanged. Additionally, we screened the mRNA expressions of 47 types of different glycosyltransferases and found that α1,6-fucosyltransferase 8 (FUT8) was the most up-regulated and contributed to strengthen the LCA binding capability to fucosylated modified ANXA2 and HSP90B1 after HCV infection. HCV infection caused the altered N-glycans profiles, increased expressions of FUT8, fucosylated ANXA2 and HSP90B1 as well as enhanced LCA binding to Huh7.5.1. Our results may lay the foundation for clarifying the role of N-glycans and facilitate the development of novel diagnostic biomarkers and therapeutic targets based on the increased FUT8, fucosylated ANXA2 and HSP90B1 after HCV infection. •Altered profiles of N-glycans in the HCVcc-infected Huh7.5.1 cells were analyzed.•Fucosylated N-glycans are significantly elevated after HCV infection.•LCA-binding glycoconjugates are the most increased in HCVcc-infected cells.•Fucosylated ANXA2/HSP90B1 are greatly increased in HCV-infected Huh7.5.1 cells.•FUT8 contributes to the elevated fucosylations of ANXA2 and HSP90B1.