[Inhibition of hepatitis B virus replication and expression by RNA interference in vitro].
OBJECTIVE
To design pSilencer3.1-H1hygro plasmid expressing short interfering RNAs (siRNA) that target HBV S gene region, and to evaluate inhibitory effect of this siRNA on HBV in vitro.


METHODS
HepG2.2.15 was used as target cell. The plasmid expressing small interfering RNA was transfected into the cultured cells via liposome metafectene, HBsAg and HBeAg were analyzed by time-resolved immunofluorometric assay, HBV DNA were analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV S-mRNA was detected by semi-quantitative RT-PCR.


RESULTS
The plasmid expressing siRNA was successfully constructed. The S region siRNAs could effectively inhibit both antigens secretion and HBV replication compared with controls, HBsAg levels decreased by 75%, 82%, 89%; HBeAg levels decreased by 32%, 38%, 43%; HBV DNA production decreased by 30%, 43%, 49%; The HBV mRNA species was reduced by 30%, 70%, 90% when transfected with 1 microg, 2 microg, 4 microg HBV S-siRNA, respectively.


CONCLUSIONS
These results demonstrate that RNAi can substantially inhibit HBV replication and the antigens expression in the infected cells. These inhibitive effect of siRNA on HBV was dose-dependent and sequence-specific.
Zhu
Cai
C
Department of Infectious Diseases, Xiangya Hospital, Central South University, Changsa 410008, China.
Fan
Xue-gong
XG
Li
Ning
N
Ying
Ruo-su
RS
Tian
Xue-fei
XF
D015967:Gene Expression Regulation, Viral; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D006801:Humans; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D014779:Virus Replication
D006514:Hepatitis B Surface Antigens; D034741:RNA, Small Interfering