Inhibition of hepatitis C virus replication by single and dual small interfering RNA using an HCV-infected cell model
Inhibition of hepatitis C virus replication by single and dual small interfering RNA using an HCV-infected cell model
复制标题
使用 HCV 感染细胞模型通过单和双小干扰 RNA 抑制丙型肝炎病毒复制
作者：
Xiaowei Xing;Su;Ji;Zhi Chen
针对丙型肝炎病毒（HCV）基因不同区域的双siRNA协同抑制HCV RNA的复制。建立HCV感染细胞模型，采用RT-PCR和Western blot检测HCV RNA和核心蛋白。设计了四种HCV特异性siRNA（siCore、siNS 3、siNS 4 B、siNS 5 B），并转染HCV感染的Huh7.5.1细胞。采用真实的时间PCR和琼脂糖凝胶电泳比较siRNA的抗病毒效果。IFNα-2b可抑制HCV在感染细胞中的复制，并呈剂量依赖性。与单一siRNA处理相比，用低剂量（0.1和10 nM）的任何两种siRNA（siCore、siNS 3和siNS 5 B）的组合处理实现了协同抑制作用（P < 0.05）。CCK-8检测结果显示siRNA对Huh7.5.1细胞无毒性作用。这些发现表明了一种有前途的治疗HCV的新方法。
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.
DOI：
10.1073/pnas.0503596102
发表时间：
2005-06-28
影响因子：
11.1
作者：
Zhong, J;Gastaminza, P;Chisari, FV
通讯作者：
Chisari, FV