Microarray analysis of gene expression in the liver of transgenic mouse model of HCV infection

Masoud Ghorbani,Catalina Soare,Turaya Naas,Francisco Diaz-Mitoma,Rashmi Kothary

doi:10.4236/abb.2012.38141

Masoud Ghorbani, Catalina Soare + Show 3 more

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https://doi.org/10.4236/abb.2012.38141

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Abstract

Background: The molecular interactions of hepatitis C virus (HCV) with hepatic tissue have yet to be completely elucidated and understood. The purpose of this study was to compare differential gene expression patterns in the livers of non-transgenic and transgenic mouse model expressing HCV structural proteins Core, Envelope 1 (E1) and Envelope 2 (E2) using complementary DNA (cDNA) microarrays. Results: Total RNA extracted from the livers of HCV transgenic and non-transgenic mice was analyzed with cDNA microarray and differentially expressed genes confirmed by real-time RT-PCR. Relative expression ratios of individual genes were determined by comparing hybridization of Cy5-labelled cDNA from transgenic mouse livers and Cy3-labelled cDNA from non-transgenic mouse livers. The spot array images were quantified using QuantArray software and the outlier spots was normalized and filtered using five different criteria. 15,297 genes were analyzed using three different analytical methods. Depending on these methods, twenty-one genes were found to be differentially expressed at a statistically significant level. From these, 6 genes had a consistent differential expression. Several genes were directly involved in lipid metabolism and lipid β-oxidation. 5-azacytidine induced gene 2 (AZ2), which is involved in the methylation of genes was down regulated in HCV transgenic mice. Altered transcript levels of these 6 genes were confirmed by real-time RT-PCR analysis. Conclusion: Interactions between HCV and hepatocytes not only involve lipid metabolism and redox balance, but this interaction may also influence DNA methylation, indicating a potential association with the development of hepatocellular carcinoma.

Highlights

The molecular interactions of hepatitis C virus (HCV) with hepatic tissue have yet to be completely elucidated and understood
More than 80% of HCV-infected individuals develop the chronic form of the disease and 20% of chronically infected patients will develop cirrhosis, and many of those will progress to hepatocellular carcinoma [3,4]
Countless efforts over the past decade have attempted to understand the pathogenesis of HCV infection, the mechanisms leading to viral persistence and disease progression are not completely understood

Summary

Introduction

The molecular interactions of hepatitis C virus (HCV) with hepatic tissue have yet to be completely elucidated and understood. The purpose of this study was to compare differential gene expression patterns in the livers of non-transgenic and transgenic mouse model expressing HCV structural proteins Core, Envelope 1 (E1) and Envelope 2 (E2) using complementary DNA (cDNA) microarrays. Results: Total RNA extracted from the livers of HCV transgenic and non-transgenic mice was analyzed with cDNA microarray and differentially expressed genes confirmed by real-time RT-PCR. Conclusion: Interactions between HCV and hepatocytes involve lipid metabolism and redox balance, but this interaction may influence DNA methylation, indicating a potential association with the development of hepatocellular carcinoma. Perhaps the most significant phenotype observed in core transgenic mice was the induction of steatosis and hepatocellular carcinoma (HCC), both of which are hallmarks of chronic infections with HCV [9,10,11]. Increased lipid peroxidation and mitochondrial DNA damage, both indicators of increased reactive oxygen species (ROS), have been noted in some

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