title: Alteration of N-glycan expression profile and glycan pattern of glycoproteins in human hepatoma cells after HCV infection.
abstract: 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.
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China.
authors.forename: Tian
authors.initials: T
authors.lastname: Xiang
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Ganglong
authors.initials: G
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Liu
authors.affiliation: University of Illinois at Urbana-Champaign, School of Molecular and Cellular Biology, Department of Microbiology, IL 61801, USA.
authors.forename: Yidan
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China.
authors.forename: Zhongxiao
authors.initials: Z
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Lu
authors.affiliation: Division of Regulatory Glycobiology, Tohoku Medical and Pharmaceutical University, 4-4-1 Komatsushima, Aobaku, Sendai, Miyagi 981-8558, Japan.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Gu
authors.affiliation: Systems Glycobiology Group, Global Research Cluster, RIKEN and RIKEN-Max Planck Joint Research Center, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Zengqi
authors.initials: Z
authors.lastname: Tan
authors.affiliation: Wuhan Institute of Biotechnology, Medical Research Institute of Wuhan University, Wuhan 430071, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xie
authors.affiliation: The Key Laboratory of Carbohydrate Chemistry & Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China. Electronic address: fengguan@jiangnan.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Guan
authors.affiliation: State Key Laboratory of Virology. Hubei province Key Laboratory of Allergy and Immune-related diseases, Medical Research Institute, Department of Immunology of Wuhan University School of Basic Medical Sciences, Wuhan 430071, China. Electronic address: zhangxiaolian@whu.edu.cn.
authors.forename: Xiao-Lian
authors.initials: XL
authors.lastname: Zhang
mesh_terms: D017306:Annexin A2 / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000821:virology; D002460:Cell Line; D005643:Fucose / Q000378:metabolism; D005647:Fucosyltransferases / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D016695:Glycosyltransferases / Q000378:metabolism; D016174:Hepacivirus / Q000472:pathogenicity; D006526:Hepatitis C / Q000378:metabolism* / Q000821:virology; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000821:virology; D008103:Liver Cirrhosis / Q000378:metabolism / Q000821:virology; D008113:Liver Neoplasms / Q000378:metabolism* / Q000821:virology; D008562:Membrane Glycoproteins / Q000378:metabolism; D037121:Plant Lectins / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*
chemical_list: D017306:Annexin A2; D006023:Glycoproteins; D037102:Lectins; D008562:Membrane Glycoproteins; D037121:Plant Lectins; D011134:Polysaccharides; C053387:endoplasmin; C033619:lentil lectin; D005643:Fucose; D016695:Glycosyltransferases; D005647:Fucosyltransferases
keywords: Glycosyltransferase expression analysis; Hepatitis C virus (HCV); Lectin microarray; Mass spectrometry; N-glycan; α1,6-fucosyltransferase 8 (FUT8)