Inhibition of HBV-DNA replication and expression by siRNA based on magnetic nanoparticles transfering in HepG2 2.2.15 cells
10.3969/j.issn.1672-7347.2010.06.002
- VernacularTitle:利用纳米和RNA干扰技术抑制HBV-DNA在HepG2 2.2.15细胞中的复制和表达
- Author:
Yan HE
;
Yongfang JIANG
;
Gufeng WANG
;
Hongyu LUO
;
Xinqiang XIAO
;
Chunming DENG
;
Kaizhong LUO
;
Xianshi SU
- Publication Type:Journal Article
- Keywords:
siRNA;
HBV;
HBV core gene;
magnetic nanoparticles
- From:
Journal of Central South University(Medical Sciences)
2010;35(6):543-548
- CountryChina
- Language:Chinese
-
Abstract:
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. Conclusion Downregulation of HBV core gene using RNAi technology and magnetic nanoparticles can potentially be used as a therapeutic strategy for Hepatitis B.
