肝细胞癌异种移植支持 HCV 复制：一种用于评估抗病毒药物的小鼠模型。
Hepatocellular carcinoma xenograft supports HCV replication: a mouse model for evaluating antivirals.
机构信息
Department of Pathology and Laboratory Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA.
出版信息
World J Gastroenterol. 2011 Jan 21;17(3):300-12. doi: 10.3748/wjg.v17.i3.300.
AIM
To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral treatment.
METHODS
We developed a stable S3-green fluorescence protein (GFP) cell line that replicated the GFP-tagged HCV sub-genomic RNA derived from a highly efficient JFH1 virus. S3-GFP replicon cell line was injected subcutaneously into γ-irradiated SCID mice. We showed that the S3-GFP replicon cell line formed human HCC xenografts in SCID mice. Cells were isolated from subcutaneous tumors and then serially passaged multiple times in SCID mice by culturing in growth medium supplemented with G-418. The mouse-adapted S3-GFP replicon cells were implanted subcutaneously and also into the liver of SCID mice via intrasplenic infusion to study the replication of HCV in the HCC xenografts. The tumor model was validated for antiviral testing after intraperitoneal injection of interferon-α (IFN-α).
RESULTS
A highly tumorigenic S3-GFP replicon cell line was developed that formed subcutaneous tumors within 2 wk and diffuse liver metastasis within 4 wk in SCID mice. Replication of HCV in the subcutaneous and liver tumors was confirmed by cell colony assay, detection of the viral RNA by ribonuclease protection assay and real-time quantitative reverse transcription polymerase chain reaction. High-level replication of HCV sub-genomic RNA in the tumor could be visualized by GFP expression using fluorescence microscopy. IFN-α cleared HCV RNA replication in the subcutaneous tumors within 2 wk and 4 wk in the liver tumor model.
CONCLUSION
A non-infectious mouse model allows us to study replication of HCV in subcutaneous and metastatic liver tumors. Clearance of HCV by IFN-α supports use of this model to test other anti-HCV drugs.
目的
建立一个用于研究丙型肝炎病毒（HCV）在小鼠体内复制和抗病毒治疗的肝细胞癌（HCC）异种移植模型。
方法
我们构建了一个稳定的 S3-绿色荧光蛋白（GFP）细胞系，该细胞系复制了源自高效 JFH1 病毒的 GFP 标记 HCV 亚基因组 RNA。将 S3-GFP 复制子细胞系皮下注射到γ射线照射的 SCID 小鼠中。我们表明，S3-GFP 复制子细胞系在 SCID 小鼠中形成了人 HCC 异种移植物。从皮下肿瘤中分离细胞，并在含有 G-418 的生长培养基中在 SCID 小鼠中连续传代多次。将适应小鼠的 S3-GFP 复制子细胞皮下植入和通过脾内输注植入 SCID 小鼠的肝脏中，以研究 HCV 在 HCC 异种移植物中的复制。通过腹腔内注射干扰素-α（IFN-α）对肿瘤模型进行抗病毒测试验证。
结果
我们开发了一个高度致瘤性的 S3-GFP 复制子细胞系，该细胞系在 2 周内形成皮下肿瘤，在 4 周内形成弥漫性肝转移。通过细胞集落测定、核糖核酸酶保护测定和实时定量逆转录聚合酶链反应检测到病毒 RNA，证实了 HCV 在皮下和肝肿瘤中的复制。使用荧光显微镜可以通过 GFP 表达可视化肿瘤中 HCV 亚基因组 RNA 的高水平复制。IFN-α 在 2 周和 4 周内清除了皮下肿瘤中的 HCV RNA 复制。
结论
非传染性小鼠模型使我们能够研究 HCV 在皮下和转移性肝肿瘤中的复制。IFN-α 清除 HCV 支持使用该模型测试其他抗 HCV 药物。
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