Inhibition of HBV-DNA replication and expression by siRNA based on magnetic nanoparticles transfering in HepG2 2.2.15 cells.
OBJECTIVE
To investigate the inhibitory effect of downregulation of hepatitis B virus (HBV) core gene (HBcAg) expression by RNA interference and magnetic nanoparticles on both HBV DNA replication and expression in vitro.


METHODS
HepG2 2.2.15 cells were transfected with U6 promoter plasmids coding for small interfering RNA (siRNA) targeting HBV core gene using magnetic nanoparticles. RT-PCR and Western blot were used to assess the mRNA and protein expression HBV core antigen. Real-time PCR was used to evaluate the suppression efficiency of HBV-DNA replication and expression; and radioimmunoassay was used for HBV surface antigen (HBsAg), core antigen (HBcAg), and e antigen (HBeAg) detection.


RESULTS
We successfully constructed nanoparticles with siRNA plasmid targeting HBV core antigen; HBcAg mRNA and HBV core antigen protein levels were significantly reduced in the transfected cells. HBV-DNA downregulation was estimated at 4-5 logs and the HBsAg and HBeAg levels were also reduced compared with the controls.


CONCLUSIONS
Downregulation of HBV core gene using RNAi technology and magnetic nanoparticles can potentially be used as a therapeutic strategy for Hepatitis B.
He
Yan
Y
Institute of Liver Diseases, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Jiang
Yongfang
Y
Wang
Gufeng
G
Luo
Hongyu
H
Xiao
Xinqiang
X
Deng
Chunming
C
Luo
Kaizhong
K
Su
Xianshi
X
D004279:DNA, Viral / Q000235:genetics*; D015536:Down-Regulation; D056945:Hep G2 Cells; D006512:Hepatitis B Core Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000502:physiology*; D006801:Humans; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D012313:RNA / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication
D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D058185:Magnetite Nanoparticles; D034741:RNA, Small Interfering; D012313:RNA