Downregulation of HBx mRNA in HepG2.2.15 cells by small interfering RNA.
OBJECTIVE
Hepatitis B virus (HBV) infection is one of the most prevalent viral infectious diseases in humans. And it is still a challenge for the development of an effective therapy for HBV infection. Recently, the progress in RNA interference (RNAi) has shed some light on the inhibition of HBV expression and replication by RNAi specific for the various genes of the HBV genome. Some prior researches suggests that the HBV x protein (HBx) plays an important role in viral transcription, cell growth, and apoptotic cell death.


METHODS
In the present study, we designed three siRNAs based on the X-protein of HBV sequences and tested their effects on the expression of HBx gene following sorting of siRNA-positive cells. The interference effect was tested in 24, 48, and 72 h. HBsAg in cultured media was assayed using western blot at various days post-transfection. The amount of HBx mRNA was quantitated by Real-time reverse-transcript PCR (RT-PCR).


RESULTS
There was a decrease in the levels of HBV mRNA and HBsAg from the the transfected cells. Among these three siRNAs, siRNA-2 was found to be the most effective at suppressing HBV gene expression.
Jiao
Jing
J
Department of Radiation Medicine, Southern Medical University, Guangzhou, Guangdong Province, PR China.
Cao
Hong
H
Chen
Xiao-Wei
XW
Zhou
Mei-Juan
MJ
Liu
Zhi-Hua
ZH
Ding
Zhen-Hua
ZH
D015153:Blotting, Western; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015534:Trans-Activators / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D054334:Viral Regulatory and Accessory Proteins; D014779:Virus Replication / Q000235:genetics
D006514:Hepatitis B Surface Antigens; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein