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Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice
Published on: September 25, 2019
Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells
Jun Liu1, Ying Guo, Cai-Fang Xue
1Department of Etiology, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
Aim:
To study the effect of siRNA expressed from DNA vector on HBV replication.
Methods:
Human U6 promoter was amplified from genomic DNA and cloned into plasmid pUC18 to construct a mammalian siRNA expression vector pUC18U6. Then oligonucleotides coding for a short hairpin RNA against HBV were cloned into pUC18U6 to form pUC18U6HBVsir which was introduced into 2.2.15 cells by using liposome-mediated transfection. 2.2.15 cells transfected by pUC18U6 and pUC18U6GFPsir which expressed siRNA against green fluorescent protein and mock-transfected 2.2.15 cells were used as controls. Concentration of HBsAg in the supernatant of the transfected cells was measured by using solid-phase radioimmunoassay.
Results:
A mammalian siRNA expression vector pUC18U6 was constructed successfully. Compared with controls, pUC18U6HBVsir which expressed siRNA against HBV decreased concentration of HBsAg significantly by 44%(P<0.05).
Conclusion:
HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.
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