Effect of vector-expressed siRNA on HBV replication in hepatoblastoma cells.
OBJECTIVE
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).


CONCLUSIONS
HBV replication in 2.2.15 cells is inhibited by siRNA expressed from the DNA vector.
Liu
Jun
J
Department of Etiology, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
Guo
Ying
Y
Xue
Cai-Fang
CF
Li
Ying-Hui
YH
Huang
Yu-Xiao
YX
Ding
Jin
J
Gong
Wei-Dong
WD
Zhao
Ya
Y
D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics*; D006509:Hepatitis B / Q000503:physiopathology / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development*; D018197:Hepatoblastoma; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication / Q000235:genetics
D034741:RNA, Small Interfering