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Combination of small interfering RNA and lamivudine on inhibition of human B virus replication in HepG2.2.15 cells.小干扰RNA与拉米夫定联合抑制人乙型肝炎病毒在HepG2.2.15细胞中的复制
World J Gastroenterol. 2007 Apr 28;13(16):2324-7. doi: 10.3748/wjg.v13.i16.2324.
2
Inhibition of Hepatitis B virus cccDNA replication by siRNA.小干扰RNA对乙型肝炎病毒共价闭合环状DNA复制的抑制作用
Biochem Biophys Res Commun. 2007 Apr 6;355(2):404-8. doi: 10.1016/j.bbrc.2007.01.163. Epub 2007 Feb 6.
3
siRNA-resistance in treated HCV replicon cells is correlated with the development of specific HCV mutations.经治疗的丙型肝炎病毒（HCV）复制子细胞中的小干扰RNA（siRNA）抗性与特定HCV突变的发生相关。
J Viral Hepat. 2006 Nov;13(11):756-61. doi: 10.1111/j.1365-2893.2006.00752.x.
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Virus Res. 2005 Sep;112(1-2):100-7. doi: 10.1016/j.virusres.2005.04.001. Epub 2005 Apr 26.
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RNA interference-mediated control of hepatitis B virus and emergence of resistant mutant.RNA干扰介导的乙肝病毒控制及耐药突变体的出现
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Clearance of hepatitis B virus from the liver of transgenic mice by short hairpin RNAs.通过短发夹RNA清除转基因小鼠肝脏中的乙型肝炎病毒
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[Potent and specific inhibition of hepatitis B virus antigen expression by RNA interference].[RNA干扰对乙型肝炎病毒抗原表达的强效特异性抑制作用]
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小干扰RNA组合对HepG2.2.15细胞中的乙型肝炎病毒共价闭合环状DNA具有更强的抑制作用。

Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells.

作者信息

Xin Xiao-Min, Li Gui-Qiu, Jin Ying-Yu, Zhuang Min, Li Di

机构信息

Department of Laboratory Diagnosis, the First Affiliated Hospital of Harbin Medical University, Heilongjiang Province, Harbin, China.

出版信息

World J Gastroenterol. 2008 Jun 28;14(24):3849-54. doi: 10.3748/wjg.14.3849.

DOI:10.3748/wjg.14.3849
PMID:18609708
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2721441/
Abstract

AIM

To observe the inhibition of hepatitis B virus (HBV) replication and expression in HepG2.2.15 cells by combination of small interfering RNAs (siRNAs).

METHODS

Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay (ELISA). Intracellular viral DNA and covalently closed circular DNA (cccDNA) were quantified by real-time polymerase chain reaction (PCR). HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR (RT-PCR).

RESULTS

siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dose-dependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantly, combination of siRNAs significantly suppressed HBV cccDNA amplification.

CONCLUSION

Combination of siRNAs mediates a stronger inhibition on viral replication and antigen expression in HepG2.2.15 cells, especially on cccDNA amplification.

摘要

目的

观察小干扰RNA（siRNA）联合应用对HepG2.2.15细胞中乙型肝炎病毒（HBV）复制和表达的抑制作用。

方法

构建重组质粒psil-HBV并转染至HepG2.2.15细胞。转染后48小时、72小时和96小时，收集培养基并收获细胞用于HBV复制检测。采用酶联免疫吸附测定（ELISA）检测细胞培养基中的HBsAg和HBeAg。通过实时聚合酶链反应（PCR）对细胞内病毒DNA和共价闭合环状DNA（cccDNA）进行定量。采用逆转录PCR（RT-PCR）对HBV病毒mRNA进行逆转录和定量。

结果

siRNA显示出显著的抗HBV作用。siRNA能够以剂量依赖的方式特异性抑制HBsAg的表达和HBV DNA的复制。此外，与单独使用每种siRNA相比，siRNA联合应用对抗原表达和病毒复制的抑制作用更强。更重要的是，siRNA联合应用显著抑制了HBV cccDNA的扩增。

结论

siRNA联合应用对HepG2.2.15细胞中的病毒复制和抗原表达具有更强的抑制作用，尤其是对cccDNA扩增的抑制作用。