Stable inhibition of hepatitis B virus expression and replication by expressed siRNA.
RNA interference might be an efficient antiviral therapy for some obstinate illness. Here, we studied the effects of hepatitis B virus (HBV)-specific 21-nt small interfering RNAs (siRNA) on HBV gene expression and replication in 2.2.15 cells. Seven vectors expressing specific hairpin siRNA driven by the RNA polymerase II-promoter were constructed and transfected into 2.2.15 cells. In the cell strain that can stably express functional siRNA, the HBV surface antigen (HBsAg) and the HBV e antigen (HBeAg) secretion into culture media was inhibited by 86% and 91%, respectively, as shown by an enzyme-linked immunosorbent assay. Immunofluorescence and Western blot indicated similar results. HBV DNA was markedly restrained by 3.28-fold, as assessed by the fluorescent quantitation PCR. Moreover, the HBV mRNA was significantly reduced by 80% based on semiquantitative RT-PCR. In conclusion, the specific siRNA can knock down the HBV gene expression and replication in vitro, and the silence effects have no relationship with interferon response.
Ren
Guang-Li
GL
The State Key Discipline and Diagnosis and Treatment Center of Infectious Diseases of Chinese People Liberation Army, Tang Du Hospital, Fourth Military Medical University, Xin Yi Road, Fang Zhi District, Xi'an 710038, China. rainewly@163.com
Bai
Xue-Fan
XF
Zhang
Yan
Y
Chen
Hong-Mei
HM
Huang
Chang-Xing
CX
Wang
Ping-Zhong
PZ
Li
Guang-Yu
GY
Zhang
Ying
Y
Lian
Jian-Qi
JQ
D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral / Q000235:genetics; D020868:Gene Silencing; D042822:Genomic Instability / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics* / Q000254:growth & development*; D018197:Hepatoblastoma / Q000821:virology*; D006801:Humans; D008113:Liver Neoplasms / Q000821:virology*; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication / Q000235:genetics*
D034741:RNA, Small Interfering