Abstract
BackgroundHepatitis C virus (HCV) core protein and nonstructural protein 4B (NS4B) are potentially oncogenic. Aberrant activation of the Wnt/β-catenin signaling pathway is closely associated with hepatocarcinogenesis. We investigated the effects of HCV type 1b core protein and NS4B on Wnt/β-catenin signaling in various liver cells, and explored the molecular mechanism underlying HCV-related hepatocarcinogenesis.ResultsCompared with the empty vector control, HCV core protein and NS4B demonstrated the following characteristics in the Huh7 cells: significantly enhanced β-catenin/Tcf-dependent transcriptional activity (F = 40.87, P < 0.01); increased nuclear translocation of β-catenin (F = 165.26, P < 0.01); upregulated nuclear β-catenin, cytoplasmic β-catenin, Wnt1, c-myc, and cyclin D1 protein expression (P < 0.01); and promoted proliferation of Huh7 cells (P < 0.01 or P < 0.05). Neither protein enhanced β-catenin/Tcf-dependent transcriptional activity in the LO2 cells (F = 0.65, P > 0.05), but they did significantly enhance Wnt3a-induced β-catenin/Tcf-dependent transcriptional activity (F = 64.25, P < 0.01), and promoted the nuclear translocation of β-catenin (F = 66.54, P < 0.01) and the Wnt3a-induced proliferation of LO2 cells (P < 0.01 or P < 0.05). Moreover, activation of the Wnt/β-catenin signaling pathway was greater with the core protein than with NS4B (P < 0.01 or P < 0.05).ConclusionsHCV core protein and NS4B directly activate the Wnt/β-catenin signaling pathway in Huh7 cells and LO2 cells induced by Wnt3a. These data suggest that HCV core protein and NS4B contribute to HCV-associated hepatocellular carcinogenesis.
Highlights
Hepatitis C virus (HCV) core protein and nonstructural protein 4B (NS4B) are potentially oncogenic
Real-time quantitative PCR (qPCR) was used to investigate the expression of HCV core and NS4B mRNAs, and western blotting was used to investigate the expression of HCV core and NS4B proteins in the stable cell lines
Western blotting revealed the presence of the core protein (21 kDa) in the Huh7-Core cells and NS4B (27 kDa) in the Huh7-NS4B cells, whereas those proteins were not detected in the Huh7-mkate2 cells (Fig. 1a-b)
Summary
Hepatitis C virus (HCV) core protein and nonstructural protein 4B (NS4B) are potentially oncogenic. We used reverse transcription–polymerase chain reaction (RT-PCR) to amplify the HCV core protein and NS4B genes from the sera of patients with chronic hepatitis C genotype 1b [17], and fused them with the mkate gene, which expresses red fluorescent protein Later, these genes were inserted into the lentiviral expression vector pLenti6.3/V5. We successfully constructed human HCC Huh and normal human liver cell line LO2 models that stably expressed HCV core protein and NS4B Based on this development, in the present study we explored the effects of HCV type 1b core protein and NS4B on the Wnt/β-catenin signaling pathway in Huh and LO2 cells, and examined the roles of those two proteins in the formation of HCV-related HCC.
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