Effects Of HCV Core Protein On The WNT/β-catenin/TCF Signaling Pathway And Apoptosis In Huh7Cells

Objective: To investigate the effects of HCVcore protein on Wnt/β-Catenin/TCFsignaling pathway and apoptosis in Huh7cells, and to study the relation betweenHCV core protein and Wnt/β-Catenin/TCF signaling pathway and apoptosispreliminary.Method:1.Huh7cells were transfected with the recombination expression vectorpcDNA3.1(-)/HCV-1b-core by lipofectamine(group pcDNA3.1(-)-C).The expressionof HCV core mRNA and protein was detected by RT-PCR and Western Blotrespectively. In huh7cells,effects of HCV core on proliferation was examined byMTT，expression of β-catenin was detected by RT-PCR and Western blot.We use dualluciferase assay system to detect TCF transcription activity,then we analyzed theeffects of HCV core on cell apoptosis by Flow cytometry and expression of Caspase-3by Western blot in huh7cells which induced by serum starvation.Huh7cellstransfected with the plasmid pcDNA3.1(-) and untreated Huh7cells set as controlexperiments.Result:1.The recombinant plasmid pcDNA3.1(-)/HCV-1b-core Huh7transientlytransfected Huh7cells expressed HCV core mRNA and related protein successfully.2.At different time point(24h,36h,48h,60h,72h),we can observed that proliferationrates of Huh7cells in group pcDNA3.1(-)-C is higher than pcDNA3.1(-) and blankcontrol group,the difference has statistical significance（P<0.01).3.For the relative expression of mRNA and protein of β-catenin in Huh7cells,Cells in group pcDNA3.1(-)-C were higher than cells in group pcDNA3.1(-)andblank control group significantly(P<0.01),while group pcDNA3.1(-) and blankcontrol group has no significant difference (P>0.05). 4.HCV core can improve pTOPFLASH relative luciferase activity significantly(P<0.01) compared with group pcDNA3.1(-) and blank control group.while each grouptransfected with pFOPFLASH relative luciferase activity has no significant difference(P>0.05).5.Compared with group pcDNA3.1(-) and blank control group,apoptosis rate ofgroup pcDNA3.1(-)-C decreased, and the difference is statistically significant(P<0.01).6.Compared to group pcDNA3.1(-) and blank control group,Huh7cells in grouppcDNA3.1(-)-C had a higher expression level of Caspase-3(P <0.01).Conclusion:1.Expression of HCV1b genotype core protein in Huh7cells can promote cellproliferation.2.In Huh7cells,HCV1b genotype core protein can upregulate expression ofβ-catenin protein and improve TCF transcription activity,suggesting that HCV coreprotein activated Wnt/β-catenin/TCF signaling pathway in Huh7cells.3.After induced by serum starvation,expression of HCV1b genotype core proteinin Huh7cells inhibited cell apoptosis.