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Hepatocellular carcinoma xenograft supports HCV replication:A mouse model for evaluating antivirals

World Journal of Gastroenterology 2011年第3期17卷 300-312页

作者： Sidhartha Hazari Henry J Hefler Partha K Chandra bret poat Feyza Gunduz Tara Ooms Tong Wu Luis A Balart Srikanta Dash Department of Pathology and Laboratory Medicine Tulane University Health Sciences Center New Orleans LA 70112 United States Department of Gastroenterology and Hepatology Tulane University Health Sciences Center New Orleans LA 70112 United States Department of Comparative Medicine Tulane University Health Sciences Center New Orleans LA 70112 United States

AIM: To develop a hepatocellular carcinoma (HCC) xenograft model for studying hepatitis C virus (HCV) replication in a mice, and antiviral ***: 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.

关键词： Hepatitis C virus Hepatocellular carcinoma Tumor xenograft SCID mouse Interferon-α Antiviral agent Virus replication

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