Biomedicine and Chemical Sciences 2(2) (2023) 131-138 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 Rana Mohammed Abdulnabi 1* , Rasha Munther Othman 1 & Hussein Katai Abdul Sada 2 1 Department of Microbiology, College of Veterinary Medicine, University of Basrah, Basrah – Iraq 2 Department of Microbiology, Al-Zahra College of Medicine, University of Basrah, Basrah – Iraq A R T I C L E I N F O A B S T R A C T Article history: 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. Copyright © 2023 Biomedicine and Chemical Sciences. Published by International Research and Publishing Academy – Pakistan, Co-published by Al-Furat Al-Awsat Technical University – Iraq. This is an open access article licensed under CC BY: (https://creativecommons.org/licenses/by/4.0) Received on: January 17, 2023 Revised on: February 5, 2023 Accepted on: February 15, 2023 Published on: April 01, 2023 Keywords: Hepatitis E virus HEV immune response HEV ORF3 HEV ORF3 plasmid Transfection 1. Introduction 1Hepatitis E virus V (HEV) outbreaks are increasingly reported. Infection with HEV can cause acute hepatitis or, in rare cases, fulminant hepatic failure, although the disease is typically sporadic and self-limiting. Throughout the second and third months of pregnancy, HEV infection is also associated with high mortality rates of around 20 percent (Wu et al., 2020). The virus is small, non-enveloped (27-34 * Corresponding author: Rana Mohammed Abdulnabi, Department of Microbiology, College of Veterinary Medicine, University of Basrah, Basrah – Iraq E-mail: ranaalkafji@gmail.com How to cite: Abdulnabi, R. M., Othman, R. M., & Sada, H. K. A. (2023). 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. Biomedicine and Chemical Sciences, 2(2), 131–138. DOI: https://doi.org/10.48112/bcs.v2i2.442 nm in diameter) and belongs to the family Hepeviridae, genus Orthohepevirus3, It has a linear, single-stranded, plus-sense RNA genome with three open reading frames: ORF1, ORF2, and ORF3 (Ledesma et al., 2019). ORF1 encodes a non-structural polyprotein associated with HEV replication (Cierniak et al., 2022), while ORF2 encodes a capsid protein involved in a virion assembly and immunogenicity (Boumaiza et al., 2021). ORF3 encodes a multifunctional phosphoprotein that suppresses host inflammation, protects virus-infected cells, and links intracellular signal transduction pathways (Maurer et al., 2022). The virus has four different genotypes but only one serotype. Genotypes 1 and 2 are restricted to human transmission, but genotypes 3 and 4 are zoonotic. There is a higher prevalence of HEV genotype 1 in young people with acute hepatitis (Wu et al. 2020). To replicate and cause pathogenicity, The HEV, like the majority of other viruses, modifies the machinery of the host cell. It would suggest that the ORF3 protein (pORF3) reduced the inflammatory response of the host cell (Chandra et al. 2011). This study aimed to analyze the effect of HEV ORF3 protein on the Content lists available at: www.journals.irapa.org/index.php/BCS/issue/view/23 Biomedicine and Chemical Sciences J o u r n a l h o m e p a g e : https://www.journals.irapa.org/index.php/BCS 10-BCS-1808-442