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Inhibition of hepatitis C virus replication by single and dual small interfering RNA using an HCV-infected cell model

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Abstract

Dual siRNA against different regions of gene in hepatitis C virus (HCV) synergistically inhibited replication of HCV RNA. An HCV-infected cell model was established, and HCV RNA and core protein were detected by RT-PCR and Western blot, respectively. Four HCV-specific siRNAs (siCore, siNS3, siNS4B, siNS5B) were designed and transfected into HCV-infected Huh7.5.1 cells. The antiviral efficacies of the siRNAs were compared using real time PCR and agarose gel electrophoresis. HCV replication in infected cells was inhibited by IFNα-2b in a dose-dependent manner. Synergistic inhibition effects were achieved with combination treatment of any two of the siRNAs (siCore, siNS3 and siNS5B) at low doses (0.1 and 10 nM), as compared to single siRNA treatment (P < 0.05). Furthermore, CCK-8 assay showed no toxicity of the siRNAs to Huh7.5.1 cells. These findings indicate a promising new therapeutic approach for treatment of HCV.

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References

  • Chang B, Lee CH, Lee JH, Lee SW (2010) Comparative analysis of intracellular inhibition of hepatitis C virus replication by small interfering RNAs. Biotechnol Lett 32:1231–1237

    Article  PubMed  CAS  Google Scholar 

  • Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K, Tuschl T (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411:494–498

    Article  PubMed  CAS  Google Scholar 

  • Hannon GJ (2002) RNA interference. Nature 418:244–251

    Article  PubMed  CAS  Google Scholar 

  • Kanda T, Steele R, Ray R, Ray RB (2007) Small interfering RNA targeted to hepatitis C virus 5′ nontranslated region exerts potent antiviral effect. J Virol 81:669–676

    Article  PubMed  CAS  Google Scholar 

  • Kato T, Furusaka A, Miyamoto M, Date T, Yasui K, Hiramoto J, Nagayama K, Tanaka T, Wakita T (2001) Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient. J Med Virol 64:334–339

    Article  PubMed  CAS  Google Scholar 

  • Kato T, Date T, Miyamoto M, Furusaka A, Tokushige K, Mizokami M, Wakita T (2003) Efficient replication of the genotype 2a hepatitis C virus subgenomic replicon. Gastroenterology 125:1808–1817

    Article  PubMed  CAS  Google Scholar 

  • Lindenbach BD, Evans MJ, Syder AJ, Wolk B, Tellinghuisen TL, Liu CC, Maruyama T, Hynes RO, Burton DR, McKeating JA, Rice CM (2005) Complete replication of hepatitis C virus in cell culture. Science 309:623–626

    Article  PubMed  CAS  Google Scholar 

  • Penin F, Dubuisson J, Rey FA, Moradpour D, Pawlotsky JM (2004) Structural biology of hepatitis C virus. Hepatology 39:5–19

    Article  PubMed  CAS  Google Scholar 

  • Prabhu R, Garry RF, Dash S (2006) Small interfering RNA targeted to stem-loop II of the 5′ untranslated region effectively inhibits expression of six HCV genotypes. Virol J 3:100

    Article  PubMed  Google Scholar 

  • Sakamoto N, Tanabe Y, Yokota T, Satoh K, Sekine-Osajima Y, Nakagawa M, Itsui Y, Tasaka M, Sakurai Y, Cheng-Hsin C, Yano M, Ohkoshi S, Aoyagi Y, Maekawa S, Enomoto N, Kohara M, Watanabe M (2008) Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA. J Gastroenterol Hepatol 23:1437–1447

    Article  PubMed  CAS  Google Scholar 

  • Shepard CW, Finelli L, Alter MJ (2005) Global epidemiology of hepatitis C virus infection. Lancet Infect Dis 5:558–567

    Article  PubMed  Google Scholar 

  • Volarevic M, Smolic R, Wu CH, Wu GY (2007) Potential role of RNAi in the treatment of HCV infection. Expert Rev Anti Infect Ther 5:823–831

    Article  PubMed  CAS  Google Scholar 

  • Wilson JA, Richardson CD (2005) Hepatitis C virus replicons escape RNA interference induced by a short interfering RNA directed against the NS5b coding region. J Virol 79:7050–7058

    Article  PubMed  CAS  Google Scholar 

  • Zhong J, Gastaminza P, Cheng G, Kapadia S, Kato T, Burton DR, Wieland SF, Uprichard SL, Wakita T, Chisari FV (2005) Robust hepatitis C virus infection in vitro. Proc Natl Acad Sci USA 102:9294–9299

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

Huh7.5.1 cells were kindly provided by Scott Forrest and Jin Zhong. This work was supported by the State S&T Projects (11th Five Year) (2008ZX10002-013).

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Authors and Affiliations

  1. State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Medical College, Zhejiang University, Hangzhou, 310003, People’s Republic of China

    Xiao-kang Xing, Su-juan Li & Zhi Chen

  2. Institute of Environmental Medicine, Medical College, Zhejiang University, Hangzhou, 310058, People’s Republic of China

    Ji-liang He

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  1. Xiao-kang Xing
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  2. Su-juan Li
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  4. Zhi Chen
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Correspondence to Zhi Chen.

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Xing, Xk., Li, Sj., He, Jl. et al. Inhibition of hepatitis C virus replication by single and dual small interfering RNA using an HCV-infected cell model. Biotechnol Lett 34, 295–301 (2012). https://doi.org/10.1007/s10529-011-0761-y

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  • DOI: https://doi.org/10.1007/s10529-011-0761-y

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