Effect of HIV-1 Rev protein and DDX3 helicase on HCV RNA replication

罗亚东; 洪国枯; 贾鸣; 郭艳; 刘明; 毛青

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Effect of HIV-1 Rev protein and DDX3 helicase on HCV RNA replication

更新时间：2026-01-09

- Effect of HIV-1 Rev protein and DDX3 helicase on HCV RNA replication
- Journal of Army Medical University Vol. 37, Issue 23, Pages: 2329-2333(2015)
- 作者机构：
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  CLC： R345;R373.9;R394.3
- Published：2015
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罗亚东,洪国枯,贾鸣,郭艳,刘明,毛青.HIV-1 Rev蛋白和人DDX3解旋酶对HCV RNA复制影响[J].陆军军医大学学报(原第三军医大学学报),2015,37(23):2329-2333.

　Luo Yadong,Hong Guoku,Jia Ming,Guo Yan,Liu Ming,Mao Qing.Effect of HIV-1 Rev protein and DDX3 helicase on HCV RNA replication[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(23):2329-2333.
罗亚东,洪国枯,贾鸣,郭艳,刘明,毛青.HIV-1 Rev蛋白和人DDX3解旋酶对HCV RNA复制影响[J].陆军军医大学学报(原第三军医大学学报),2015,37(23):2329-2333. DOI：

　Luo Yadong,Hong Guoku,Jia Ming,Guo Yan,Liu Ming,Mao Qing.Effect of HIV-1 Rev protein and DDX3 helicase on HCV RNA replication[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(23):2329-2333. DOI：

摘要

目的研究人类免疫缺陷病毒Ⅰ型（HIV-1）病毒颗粒蛋白表达调节因子（regulator of virion protein expression

Rev）和DEAD-box蛋白家族ATP依赖的RNA解旋酶3（DDX3）对HCV RNA复制的影响。方法利用HCV表达质粒pCDNA3.1-JFH1转染Huh7细胞以构建稳定的HCV细胞复制模型，另构建pCDNA3.1-Rev-Flag-mCherry和pCDNA3.1-DDX3-YFP-Flag的过表达载体，通过PCR、Western blot和核苷酸测序等方法鉴定载体构建成功后，将实验分为2组对照实验，对照组均只将HCV复制质粒转染Huh7细胞：1组实验组为DDX3和HCV RNA复制质粒共转染入Huh7细胞； 2组的实验组为Rev、DDX3和HCV RNA复制质粒共转染入Huh7细胞。最后通过qPCR定量测定各组HCV RNA复制水平并进行比较。结果转染后48 h测各组HCV RNA的复制水平，DDX3+HCV RNA复制质粒组中HCV RNA的水平（1.09×10

±0.18×10

）明显高于单独转染HCV RNA复制质粒组(4.79×10

±1.24×10

)（P=0.03）；而Rev+DDX3+HCV RNA复制质粒组中HCV RNA的水平(1.74×10

±0.40×10

)明显低于单独转染HCV RNA复制质粒组（4.17×10

±0.46×10

）（ P0.001）。结论成功构建Rev蛋白和DDX3的过表达载体，并初步证明人DDX3解旋酶可促进HCV RNA的复制，而Rev蛋白和DDX3共同作用可抑制HCV RNA的复制。

Abstract

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

vs (4.79±1.24)×10

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

vs (4.17±0.46)×10

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.

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