pr5008703_si_002.xls (1.06 MB)
Quantitative Proteomic Analysis of Exosome Protein Content Changes Induced by Hepatitis B Virus in Huh‑7 Cells Using SILAC Labeling and LC–MS/MS
dataset
posted on 2014-12-05, 00:00 authored by Xue Zhao, Yanxin Wu, Jinlin Duan, Yanchun Ma, Zhongliang Shen, Lili Wei, Xiaoxian Cui, Junqi Zhang, Youhua Xie, Jing LiuHepatitis B virus
(HBV) infection could cause hepatitis, liver
cirrhosis, and hepatocellular carcinoma. HBV-mediated pathogenesis
is only partially understood, but X protein (HBx) reportedly possesses
oncogenic potential. Exosomes are small membrane vesicles with diverse
functions released by various cells including hepatocytes, and HBV
harnesses cellular exosome biogenesis and export machineries for virion
morphogenesis and secretion. Therefore, HBV infection might cause
changes in exosome contents with functional implications for both
virus and host. In this work, exosome protein content changes induced
by HBV and HBx were quantitatively analyzed by SILAC/LC–MS/MS.
Exosomes prepared from SILAC-labeled hepatoma cell line Huh-7 transfected
with HBx, wildtype, or HBx-null HBV replicon plasmids were analyzed
by LC–MS/MS. Systematic analyses of MS data and confirmatory
immunoblotting showed that HBx overexpression and HBV, with or without
HBx, replication in Huh-7 cells indeed caused marked and specific
changes in exosome protein contents. Furthermore, specific changes
in protein contents were also detected in exosomes purified from HBV-infected
patients’ sera compared with control sera negative for HBV
markers. These results illustrate a new aspect of interactions between
HBV and the host and provide the foundation for future research into
roles played by exosomes in HBV infection and pathogenesis.