The Effect Of Hepatitis E Virus Open Reading Frame 3 Protein on The Expression Levels of IFN-β,IL6,TNF-α and RANTES in SW480 Colon Cancer Cells

Abstract

The Hepatitis E virus (HEV) could induce chronic hepatitis and liver failure with high mortality through unclear mechanisms. The purpose of this study was to determine the effect of (recombinant fusion protein, ORF3 of genotype 1 HEV) on the expression levels of cytokine or chemokine genes in colon cancer cells (SW480) using a quantitative assay of “SYBR green" real-time PCR. An ORF3 protein expression system (Orf3-GFP plasmid) was synthesized in the company of GenScript (USA) and used as an expression vector, SW480 cells were infected by (Orf3-GFP plasmid) or GFP- control vector. Result of this study was approved after assessing the expression level of the ORF3 gene in ORF3 expressing cells and in control groups using ORF3 specific primer to ensure that the transfection was performed successfully, The expression levels of three genes (IFN-β, IL-6, and TNF-α) were estimated in ORF3-expressing cells using specific primers for all genes. In addition, the role of HEV Orf3 in upregulating RANTES expression levels was revealed. In conclusion: In SW480 cells, this study found that the ORF3 protein of genotype 1 HEV is capable of regulating the expression of multiple immune response genes including Interferon- β (IFN-β), Interleukin-6 (IL-6) Tumor necrosis factor-α (TNF-α) and regulated on activation, normal T cell expressed and secreted (RANTES) which may enhance viral replication and pathogenicity.