Objective      To determine the effect of the regulator of HIV-1 virion protein expression (Rev) and a DEAD box RNA helicase (DDX3) on HCV RNA replication.       Methods      pCDNA3.1-Re-Flag-mCherry and pCDNA3.1-DDX3-YFP-Flag were constructed and identified by PCR

Western blotting and

DNA sequencing. Then the Huh7 cells were divided into 2 groups

the control group (only HCV RNA replication plasmid was transfected into Huh7 cells) and the experimental group (DDX3 and HCV RNA replication plasmids were co-transfected into Huh7 cells). qPCR was used to detect the level of HCV RNA for the effects of DDX3 and HCV RNA replication plasmids.       Results      In 48 h after transfection

the expression level of HCV RNA was significantly higher in DDX3+HCV RNA replication plasmid group than HCV RNA plasmid group [(1.09±0.18)×10

7

vs (4.79±1.24)×10

6

P=0.03

]

. But the expression level of HCV RNA was significantly lower in Rev+DDX3+HCV RNA replication plasmid group than that of HCV RNA replication HCV RNA plasmid group [(1.74±0.40)×10

7

vs (4.17±0.46)×10

6

P0.001

]

.       Conclusion      The over-expression vector of DDX3 and Rev is successfully constructed

and DDX3 can accelerate the replication of HCV RNA

but Rev combined with DDX3 can inhibit the replication of HCV in vitro.