Hazari S, Hefler HJ, Chandra PK, Poat B, Gunduz F, Ooms T, Wu T, Balart LA, Dash S. Hepatocellular carcinoma xenograft supports HCV replication: A mouse model for evaluating antivirals. World J Gastroenterol 2011; 17(3): 300-312 [PMID: 21253388 DOI: 10.3748/wjg.v17.i3.300]
Corresponding Author of This Article
Srikanta Dash, PhD, Professor, Department of Pathology and Laboratory Medicine and Gastroenterology and Hepatology, Tulane University Health Sciences Center, 1430 Tulane Ave., SL-79, New Orleans, LA 70112, United States. sdash@tulane.edu
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World J Gastroenterol. Jan 21, 2011; 17(3): 300-312 Published online Jan 21, 2011. doi: 10.3748/wjg.v17.i3.300
Hepatocellular carcinoma xenograft supports HCV replication: A mouse model for evaluating antivirals
Sidhartha Hazari, Henry J Hefler, Partha K Chandra, Bret Poat, Feyza Gunduz, Tara Ooms, Tong Wu, Luis A Balart, Srikanta Dash
Sidhartha Hazari, Partha K Chandra, Bret Poat, Feyza Gunduz, Tong Wu, Srikanta Dash, Department of Pathology and Laboratory Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, United States
Henry J Hefler, Feyza Gunduz, Luis A Balart, Srikanta Dash, Department of Gastroenterology and Hepatology, Tulane University Health Sciences Center, New Orleans, LA 70112, United States
Tara Ooms, Department of Comparative Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, United States
Author contributions: Hazari S performed the majority of the experiments, acquisition and analysis of data, and participated in design of the study; Hefler HJ performed some animal experiments and helped in the antiviral studies; Chandra PK performed the real time RT-PCR and RPA; Poat B and Gunduz F performed biochemical assays and tumor cell inoculation of SCID mice; Ooms T performed the intrasplenic injection of S3-GFP cells into SCID-NOD mice and gave valuable suggestions on animal handling and care during the whole study; Wu T performed the histological evaluation of subcutaneous and liver tumors, and took all the histological images; Balart LA provided valuable suggestions and edited the manuscript; Dash S conceptualized and designed the study, evaluated the overall results, and wrote the manuscript.
Supported by Funds received from the National Cancer Institute (CA127481, CA129776), Geyer Foundation, New York, Louisiana Cancer Research Consortium and Tulane Cancer Center
Correspondence to: Srikanta Dash, PhD, Professor, Department of Pathology and Laboratory Medicine and Gastroenterology and Hepatology, Tulane University Health Sciences Center, 1430 Tulane Ave., SL-79, New Orleans, LA 70112, United States. sdash@tulane.edu
Telephone: +1-504-9882519 Fax: +1-504-9887389
Received: June 9, 2010 Revised: August 3, 2010 Accepted: August 10, 2010 Published online: January 21, 2011
Abstract
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.
