Inhibition of multiple gene expression and virus replication of HBV by stable RNA interference in 2.2.15 cells.
OBJECTIVE
Hepatitis B virus (HBV) infection is a world-wide health problem. Recent studies have demonstrated the efficacy of RNA interference (RNAi) against HBV replication at cell culture and animal levels using transient transfection. The present study was to determine whether the stable transfection of short hairpin RNA (shRNA)-producing vector could achieve potent and sustained inhibition of the HBV replication in 2.2.15 cells.


METHODS
shRNA-producing vector against HBV and the empty vector were stably transfected into the 2.2.15 cells respectively. A series of experiments were performed in the producing stable lines to determine the changes of viral protein expression and replication.


RESULTS
The HBV protein expression and viral replication were suppressed dramatically and stably by the integrated shRNA-producing vectors. Most importantly, this suppression effect persists after 30 passages.


CONCLUSIONS
Our data provided the possibility of continuous and stable inhibition of HBV protein expression and replication in patients using RNAi, suggesting a potential clinical application of this novel approach. Furthermore, the established stable transfected cell lines provided a good platform for understanding the mechanism of anti-HBV by RNAi.
Ren
Xiangrong
X
State Key Laboratory of Biocontrol and Department of Biochemistry, College of Life Sciences, Sun Yat-sen Zhongshan University, Guangzhou 510275, People's Republic of China [corrected]
Luo
Guangbin
G
Xie
Zhenhui
Z
Zhou
Lingjun
L
Kong
Xiangping
X
Xu
Anlong
A
D015152:Blotting, Northern; D015139:Blotting, Southern; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D015967:Gene Expression Regulation, Viral / Q000187:drug effects / Q000502:physiology*; D005822:Genetic Vectors / Q000494:pharmacology; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000235:genetics; D006513:Hepatitis B e Antigens / Q000032:analysis / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology*; D018197:Hepatoblastoma / Q000737:chemistry / Q000235:genetics / Q000473:pathology / Q000821:virology; D006801:Humans; D007150:Immunohistochemistry; D008113:Liver Neoplasms / Q000737:chemistry / Q000235:genetics / Q000473:pathology / Q000821:virology; D034622:RNA Interference / Q000502:physiology*; D020871:RNA Stability; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects / Q000502:physiology*
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D034741:RNA, Small Interfering; D012367:RNA, Viral