Hepatitis C virus (HCV) is a positive‐sense, single‐stranded RNA virus that causes chronic hepatitis and hepatocellular carcinoma. Cellular microRNAs (miRNAs) directly modulate the viral infectivity and indirectly through targeting virus‐related host factors. They play an essential role in the progression of different stages of HCV infection. The roles of miR‐196 family in HCV infection and hepatocellular carcinoma progression remain poorly understood. Using ViTa databases, miR‐196a as a high‐score miRNA targeting the NS₅A region of HCV genome was selected. Using dual luciferase assay and an established cell‐cultured HCV (HCVcc) system, the effect of miR‐196a on HCV genome was assessed. In silico analysis demonstrated the significant role of miR‐196a in the downregulation of HCV replication. Using dual luciferase assay, the liver‐specific miR‐196a and NS₅A gene binding was confirmed. To assess the experimental role of miR‐196a, an HCVcc system was established in the Huh 7.5 cell lines. The HCV‐RNA 1b derived from an infected patient was transfected into Huh 7.5 cells containing miR‐196a lentiviral vectors (Huh 7.5/miR‐196a), mocks (Huh 7.5/mock vector), and naïve Huh 7.5 cells. The rate of reduction of the HCV genome replication was assessed using relative real‐time PCR assay. These results represent miR‐196a overexpression and its roles in regulating HCV genome replication. However, miR‐196a may inhibit HCV replication and accelerate the early stages of apoptosis. Overexpression of miR‐196a in Huh 7.5 replicon cell is a potential new strategy to prevent hepatitis C infection. The results of this study suggest that miR‐196a directly downregulates HCV replication and may serve as a new antiviral therapy.

中文翻译：

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使用慢病毒载体通过 miR-196a 下调丙型肝炎病毒复制

丙型肝炎病毒 (HCV) 是一种正链单链 RNA 病毒，可导致慢性肝炎和肝细胞癌。细胞微小RNA（miRNA）直接调节病毒感染性，并通过靶向病毒相关宿主因子间接调节。它们在 HCV 感染的不同阶段的进展中起着至关重要的作用。miR-196 家族在 HCV 感染和肝细胞癌进展中的作用仍然知之甚少。使用 ViTa 数据库，选择 miR-196a 作为靶向HCV 基因组NS ₅ A 区域的高分 miRNA 。使用双荧光素酶测定和已建立的细胞培养 HCV (HCVcc) 系统，评估了 miR-196a 对 HCV 基因组的影响。电脑模拟分析表明 miR-196a 在下调 HCV 复制中的重要作用。使用双荧光素酶测定，肝脏特异性 miR-196a 和NS ₅ A基因结合得到证实。为了评估 miR-196a 的实验作用，在 Huh 7.5 细胞系中建立了 HCVcc 系统。来自受感染患者的 HCV-RNA 1b 被转染到含有 miR-196a 慢病毒载体（Huh 7.5/miR-196a）、模拟物（Huh 7.5/模拟载体）和初始 Huh 7.5 细胞的 Huh 7.5 细胞中。HCV 基因组复制的减少率使用相对实时 PCR 测定进行评估。这些结果代表了 miR-196a 过表达及其在调节 HCV 基因组复制中的作用。然而，miR-196a 可能抑制 HCV 复制并加速细胞凋亡的早期阶段。在 Huh 7.5 复制子细胞中过表达 miR-196a 是预防丙型肝炎感染的潜在新策略。