Abstract

THE IMPACT OF HCV KNOCKDOWN ON STAT3 AND ADI1 EXPRESSION IN HUMAN CON1 CELLS

*Shaymaa M. M. Yahya, PhD

Abstract

Hepatitis C virus (HCV) is one of the leading causes of chronic liver disease besides, it is the main causes of liver-related morbidity and mortality. There are many host genes modulate viral infection and are an underappreciated target for antiviral therapy. From the important host genes playing role in HCV infection are STAT3 and ADI1genes. In this study, siRNA targeting the 5'HCV (siHCV) was designed using siVIRUS software. This siRNA Human Con1 cells (Huh-7 expressing HCV con1 I377/NS3-3’ replicon). Cells were transfected with the siHCV siRNAs and the all star negative control at 10nM concentrations. RNA was isolated using Qiazol buffer (Qiagen, USA) according to manufacturer instruction. RNA was subsequently cleaned up using RNAeasy mini Kit (Qiagen, USA). HCV, STAT3, and ADI1 copy numbers were quantified using QuantiFast Sybergreen RT-PCR. The copy numbers were normalized to the house keeping beta actin gen. Con1 cell transfection with 10nM siHCV significantly reduced the viral replication as compared to negative control treatment. Consequently, HCV knockdown resulted in significant reduction in both STAT3 and ADI1 gene expression as compared to negative control treatment. The results from this study revealed the strong relation between STAT3 and ADI1 host genes and HCV replication. Consequently, they could be used as potential targets in the fight against HCV infection.

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