Anti-apoptotic effect of HCV core gene of genotype 3a in Huh-7 cell line

Shah Jahan,Saba Khaliq,Sajida Hassan,Waqar Ahmad,Usman A Ashfaq,Bushra Ijaz,Muhammad Hassan Siddiqi

doi:10.1186/1743-422x-8-522

Abstract

BackgroundHepatitis C virus (HCV) Core protein regulates multiple signaling pathways and alters cellular genes expression responsible for HCV induced pathogenesis leading to hepatocellular carcinoma (HCC). Prevalence of HCV genotype 3a associated HCC is higher in Pakistan as compare to the rest of world; however the molecular mechanism behind this is still unclear. This study has been designed to evaluate the effect of HCV core 3a on apoptosis and cell proliferation which are involved in HCCMethodologyWe examined the in vitro effect of HCV Core protein of genotype 3a and 1a on cellular genes involved in apoptosis by Real time PCR in liver cell line (Huh-7). We analyzed the effect of HCV core of genotype 1a and 3a on cell proliferation by MTT assay and on phosphrylation of Akt by western blotting in Huh-7 cells.ResultsThe HCV 3a Core down regulates the gene expression of Caspases (3, 8, 9 and 10), Cyto C and p53 which are involved in apoptosis. Moreover, HCV 3a Core gene showed stronger effect in regulating protein level of p-Akt as compared to HCV 1a Core accompanied by enhanced cell proliferation in Huh-7 cell line.ConclusionFrom the current study it has been concluded that reduced expression of cellular genes involved in apoptosis, increased p-Akt (cell survival gene) and enhanced cell proliferation in response to HCV 3a core confirms anti apoptotic effect of HCV 3a Core gene in Huh-7 that may lead to HCC.

Highlights

Hepatitis C virus (HCV) causes acute and chronic hepatitis which can lead to Hepatocellular carcinoma (HCC) in a significant number of patients via induction of oxidative stress, steatosis, insulin resistance, fibrosis and liver cirrhosis [1,2]
The HCV 3a Core down regulates the gene expression of Caspases (3, 8, 9 and 10), Cytochrome C (Cyto C) and p53 which are involved in apoptosis
From the current study it has been concluded that reduced expression of cellular genes involved in apoptosis, increased p-Akt and enhanced cell proliferation in response to HCV 3a core confirms anti apoptotic effect of HCV 3a Core gene in Huh-7 that may lead to hepatocellular carcinoma (HCC)

Summary

Introduction

Hepatitis C virus (HCV) causes acute and chronic hepatitis which can lead to Hepatocellular carcinoma (HCC) in a significant number of patients via induction of oxidative stress, steatosis, insulin resistance, fibrosis and liver cirrhosis [1,2]. There are increasing evidences suggesting that liver cell damage in chronic HCV infection is mediated by apoptosis [11]. The impact of apoptosis in chronic HCV infection is not well understood It may be harmful by triggering liver fibrosis, or essential in interferon (IFN) induced HCV elimination. For virtually all HCV proteins, pro- and anti-apoptotic effects have been described, especially for the Core and E2 protein [13]. HCV Core has pro- and anti-apoptotic effects in death ligand e.g., TNF-a and CD95 ligand mediated hepatocyte apoptosis has been described in a hepatoma cell line [14,15]. Several studies demonstrated binding of the HCV Core protein to p53, either inhibiting or activating p53 following with anti- or pro-apoptotic effects [17,18]. This study has been designed to evaluate the effect of HCV core 3a on apoptosis and cell proliferation which are involved in HCC

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Results

Conclusion