摘要
目的探讨通过载体在体外培养的细胞中表达发夹状siRNAs对HBV基因表达的抑制作用。方法构建表达针对HBV S基因mRNA的发夹状siRNAs的质粒pSilencer 2.1-U6-S,并与ayw亚型HBV全基因组表达质粒共转染体外培养的HepG2215细胞,用半定量RT-PCR分析目标mRNA表达丰度的变化,用ELISA法观察HBsAg表达的变化。结果siRNA处理组细胞上清中HbsAg和HBeAg含量分别比空白对照组降低了63.4%和68.0%,细胞中的2.1kb的信使RNA降低了75.2%。结论siRNA在体外培养的细胞中能有效地抑制HBV基因的表达。
Objective To study the effect of hairpin siRNAs which expressed by constructed plasmid on hepatitis B surface antigen and hepatitis B e antigen expression in vitro. Methods Constructed plasmid pSilencer 2, 1-U6-S that express hairpin siRNAs targeted to HBV S gene was co-transfected with ayw HBV expression plasmid into HepG2215 cells. ELISA and reverse transcript （RT）-PCR were used to detected the change of HBV 2.4 kb RNA, HBsAg and HBeAg in HepG2215 cells before and after treatment with siRNA. Results After treatment with siRNA, the level of 2. 1 kb RNA was reduced by 75.2% and HBsAg and HBeAg were also reduced by 63.4% and 68.0%, respectively. Conclusion siRNA can effectively inhibit replication of HBV in vitro.
出处
《实用肝脏病杂志》
CAS
2006年第4期203-205,共3页
Journal of Practical Hepatology
