Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression / 浙江大学学报(英文版)(B辑:生物医学和生物技术)
Journal of Zhejiang University. Science. B
;
(12): 236-241, 2005.
Article
in English
| WPRIM
| ID: wpr-316345
ABSTRACT
<p><b>OBJECTIVES</b>To evaluate the inhibitory effect mediated by combination of small interfering RNAs (siRNAs) targeting different sites of hepatitis B virus (HBV) transcripts on the viral replication and antigen expression in vitro.</p><p><b>METHODS</b>(1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region of HBV genome were designed and chemically synthesized. (2) HBV-producing HepG2.2.15 cells were treated with or without siRNAs for 72 h. (3) HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay. (4) Intracellular viral DNA was quantified by real-time PCR (Polymerase Chain Reaction). (5) HBV viral mRNA was reverse transcribed and quantified by real-time PCR. (6) The change of cell cycle and apoptosis was determined by flow cytometry.</p><p><b>RESULTS</b>Our data demonstrated that synthetic small interfering RNAs (siRNAs) targeting S and PreC gene could efficiently and specifically inhibit HBV replication and antigen expression. The expression of HBsAg and HBeAg and the replication of HBV could be specifically inhibited in a dose-dependent manner by siRNAs. Furthermore, our results showed that the combination of siRNAs targeting various regions could inhibit HBV replication and antigen expression in a more efficient way than the use of single siRNA at the same final concentration. No apoptotic change was observed in the cell after siRNA treatment.</p><p><b>CONCLUSION</b>Our results demonstrated that siRNAs exerted robust and specific inhibition on HBV replication and antigen expression in a cell culture system and combination of siRNAs targeting different regions exhibited more potency.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Virus Replication
/
DNA, Viral
/
Gene Expression Regulation, Viral
/
Cell Cycle
/
Hepatitis B virus
/
Apoptosis
/
RNA, Small Interfering
/
Cell Line, Tumor
/
Flow Cytometry
Limits:
Humans
Language:
English
Journal:
Journal of Zhejiang University. Science. B
Year:
2005
Type:
Article
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