State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Medical College, Zhejiang University, Hangzhou, 310003, People's Republic of China.
Biotechnol Lett. 2012 Feb;34(2):295-301. doi: 10.1007/s10529-011-0761-y. Epub 2011 Oct 5.
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.
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。建立了 HCV 感染细胞模型，分别通过 RT-PCR 和 Western blot 检测 HCV RNA 和核心蛋白。设计并转染了四种 HCV 特异性 siRNA（siCore、siNS3、siNS4B 和 siNS5B）进入 HCV 感染的 Huh7.5.1 细胞。通过实时 PCR 和琼脂糖凝胶电泳比较了 siRNA 的抗病毒功效。IFNα-2b 以剂量依赖性方式抑制感染细胞中的 HCV 复制。与单独使用 siRNA 治疗相比，低剂量（0.1 和 10 nM）联合使用两种 siRNA（siCore、siNS3 和 siNS5B）可达到协同抑制作用（P < 0.05）。此外，CCK-8 测定显示 siRNA 对 Huh7.5.1 细胞没有毒性。这些发现表明针对 HCV 的一种有前途的新治疗方法。
