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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 supatant of the transfected cells was measured by using solid-phase radioimmunoassay.RESULTS: A mammalian siRNA expression vector pUC18U6was 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.