Effect of prohibitin on hepatitis C virus replication / 临床肝胆病杂志

ABSTRACT

ObjectiveTo investigate the effect of prohibitin (PHB) on hepatitis C virus (HCV) replication. MethodsHuman hepatocarcinoma Huh-7.5 cells were transfected with the full-length genome HCV RNA in vitro to establish the full-length genome cell model of HCV, and the supernatants were collected 24, 48, 72, and 96 hours later to measure HCV copies. The indirect immunofluorescence assay was used to measure the expression of HCV core proteins, and a transmission electron microscope was used to observe the changes in the ultrastructure of HCV-infected Huh-7.5 cells and identify the full-length genome cell model of HCV. Quantitative real-time PCR and Western Blot were used to measure the expression of PHB in HCV-infected Huh-7.5 cells, and the RNA interference technique was used to measure the replication of HCV RNA. The t-test was used for comparison between groups. ResultsThe results of identification showed that the HCV full-length genome cell model was successfully established. At 24 and 48 hours after Huh-7.5 cells were transfected with HCV RNA, the mRNA expression level of PHB was 13.41±1.35 and 16.45±1.76, respectively, which differed significantly from that in the control group (101±057 and 101±087，t=29540 and 31361，both P＜0.01), and the 24- and 48-hour transfection groups showed a significantly higher expression level of PHB than the control group (both P＜0.01). At 24 and 48 hours after Huh-7.5 cells were transfected with the RNA interference plasmids of PHB, the level of HCV RNA was 64.32±5.49 and 84.45±7.06, respectively, significantly higher than that in the control group (shRNA-control group; 10.52±1.57 and 16.34±2.97, t=29538 and 25908，all P＜0.01). ConclusionIn HCV-infected Huh-7.5 cells, the increased expression of PHB is closely related to HCV RNA infection, and to a certain extent, PHB can inhibit the replication of HCV RNA.