Combination of small interfering RNAs mediates greater inhibition of human hepatitis B virus replication and antigen expression

• Published in: Journal of Zhejiang University. B. Science Vol. 6; no. 4; pp. 236 - 241
• Main Author: 陈喆 许则丰 叶景佳 姚航平 郑树 丁佳逸
• Format: Journal Article
• Language: English
• Published: China Cancer Institute, Second affiliated hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China%Immunology Institute, School of Medicine, Zhejiang University, Hangzhou 310031, China 01.04.2005; Zhejiang University Press
• Subjects: Apoptosis; Biomedicine; Cell Cycle; Cell Line, Tumor; DNA, Viral - biosynthesis; Flow Cytometry; Gene Expression Regulation, Viral - genetics; Hepatitis B e Antigens - metabolism; Hepatitis B Surface Antigens - metabolism; Hepatitis B virus - genetics; Hepatitis B virus - physiology; Humans; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; RNAs化合物; Virus Replication - genetics; 乙肝病毒; 免疫抗原; 病毒复制; HBV replication; Antigen expression; Hepatitis B virus; Combination of siRNAs
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.2005.B0236

Objectives: 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. Methods: (1) Seven siRNAs targeting surface (S), polymerase (P) or precore (PreC) region ofHBV 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. Results: 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. Conclusion: 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.