428] MICROARRAY ANALYSIS OF LIVER CELLS CONTAINING A FULL-LENGHT HEPATITIS C VIRUS REPLICON

Abstract

Introduction: Infection with hepatitis C virus (HCV) represents the major cause of liver disease, affecting more than 170 million individuals worldwide. Several data indicate that HCV replication can influence the progression and the severity of liver diseases by a modulation of cellular gene expression. Methods: To evaluate HCV-induced modification in the global expression profile, we compare gene expression of the human hepatoma cell line 2 1-5 carrying the full-length HCV replicon with the parental cell line Huh7 and the cured replicon cells in which the HCV replicon was removed by IFN-a treatment. Gene expression was examined using Applied Biosystems Human Genome Survey Arrays containing 3 1,700, 60-mer oligonucleotides probes representing a set of27,868 individual human genes and more than 1,000 control probes. The study was designed to analyze gene expression for three biological replicates for each cell line and two technical replicates for each biological replicate, for a total of 18 microarrays. The fold change of HCV replicon vs. parental or cured cell lines was analyzed by filtering the dataset of normalized signals (using p value 10.01 and 3) and performing ANOVA statistical analysis. PANTHER Classification system (Celera Genomics) was used to interpret gene expression data obtained comparing HCV replicon vs parental or cured cells. Alteration of specific genes was confirmed by Real-time PCR. Results and Discussion: Data obtained from this study indicated that HCV replication results in a specific and significant alteration of genes involved in cellular process such as endoplasmic reticulum (ER) and oxidative stress, apoptosis, lipid metabolism, immune defense and cell cycle which are relevant for HCV pathogenesis. Virul Hepatitis linit,