Abstract
Chronic hepatitis C virus (HCV) infection is one of the leading causes of severe hepatitis. The molecular mechanisms underlying HCV replication and pathogenesis remain unclear. The development of the subgenome replicon model system significantly enhanced study of HCV. However, the permissiveness of the HCV subgenome replicon greatly differs among different hepatoma cell lines. Proteomic analysis of different permissive cell lines might provide new clues in understanding HCV replication. In this study, to detect potential candidates that might account for the differences in HCV replication. Label-free and iTRAQ labeling were used to analyze the differentially expressed protein profiles between Huh7.5.1 wt and HepG2 cells. A total of 4919 proteins were quantified in which 114 proteins were commonly identified as differentially expressed by both quantitative methods. A total of 37 differential proteins were validated by qRT-PCR. The differential expression of Glutathione S-transferase P (GSTP1), Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), carboxylesterase 1 (CES1), vimentin, Proteasome activator complex subunit1 (PSME1), and Cathepsin B (CTSB) were verified by western blot. And over-expression of CTSB or knock-down of vimentin induced significant changes to HCV RNA levels. Additionally, we demonstrated that CTSB was able to inhibit HCV replication and viral protein translation. These results highlight the potential role of CTSB and vimentin in virus replication.
Highlights
Hepatitis C virus (HCV) is a positive-stranded RNA virus that causes acute and chronic hepatitis
The first is a label-free method based on peptide peak area, whereas the second is a labeled method based on iTRAQ labeling
Global characterization of the host cellular response to HCV has been developed using liquid chromatographic (LC) separations coupled with mass spectrometry (MS) for proteome analysis to identify potential gene markers of HCV-associated liver disease [42]
Summary
Hepatitis C virus (HCV) is a positive-stranded RNA virus that causes acute and chronic hepatitis. Kato et al developed an HCV genotype 2a replicon (JFH-1) that replicates efficiently in Huh cells and other human hepatocyte-derived cells lines (HepG2 and IMY-N9) [7,8,9] and nonhepatic cells lines (HeLa and HEK293) [10, 11] without adaptive mutations. These cell lines can uptake the HCV subgenomic replicon, the efficiency of replication in cells differs significantly because of host cell permissiveness. There is still no evidence to support robust replication of the HCV genotype 1 subgenomic replicon or 2a genomic replicon in HepG2 cells without the addition of external factors
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