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
DOI:
https://doi.org/10.48112/bcs.v2i2.442Abstract
Abstract Views: 104
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.
Keywords:
Hepatitis E virus, HEV immune response, HEV ORF3 plasmid, transfection, HEV ORF3
Metrics
References
Abravanel, F., Sandres-Saune, K., Lhomme, S., Dubois, M., Mansuy, J. M., & Izopet, J. (2012). Genotype 3 diversity and quantification of hepatitis E virus RNA. Journal of clinical microbiology, 50(3), 897-902. https://doi.org/10.1128/JCM.05942-11
An, I., Harman, M., & Ibiloglu, I. (2018). Case Report Bullous Hemorrhagic Dermatosis Induced by Enoxaparin. Indian Dermatol. Online J, 9, 347-350.
Aoki, Y., Sugiyama, M., Murata, K., Yoshio, S., Kurosaki, M., Hashimoto, S., ... & Kanto, T. (2015). Association of serum IFN-λ 3 with inflammatory and fibrosis markers in patients with chronic hepatitis C virus infection. Journal of gastroenterology, 50, 894-902. https://doi.org/10.1007/s00535-014-1023-2
Atwan, Z. (2016). Promyelocytic Leukaemia Protein Isoform II and Mild Heat Stress Compromise Human Adenovirus Type 5 Gene Expression (Doctoral dissertation, University of Warwick).
Boumaiza, M., Trabelsi, K., Choucha, Z., Akrouti, I., Leone, S., Picone, D., & Kallel, H. (2021). Production and characterization of a fusion form of hepatitis E virus t ORF2 capsid protein in Escherichia coli. Preparative Biochemistry & Biotechnology, 51(6), 562-569. https://doi.org/10.1080/10826068.2020.1836656
Chandra, V., Holla, P., Ghosh, D., Chakrabarti, D., Padigaru, M., & Jameel, S. (2011). The hepatitis E virus ORF3 protein regulates the expression of liver-specific genes by modulating localization of hepatocyte nuclear factor 4. PloS one, 6(7), e22412. https://doi.org/10.1371/journal.pone.0022412
Cierniak, F., Ulrich, R. G., Groschup, M. H., & Eiden, M. (2022). A Modular Hepatitis E Virus Replicon System for Studies on the Role of ORF1-Encoded Polyprotein Domains. Pathogens, 11(3), 355. https://doi.org/10.3390/pathogens11030355
Devhare, P. B., Chatterjee, S. N., Arankalle, V. A., & Lole, K. S. (2013). Analysis of antiviral response in human epithelial cells infected with hepatitis E virus. PloS one, 8(5), e63793. https://doi.org/10.1371/journal.pone.0063793
Fitzgerald, K. A., & Kagan, J. C. (2020). Toll-like receptors and the control of immunity. Cell, 180(6), 1044-1066. https://doi.org/10.1016/j.cell.2020.02.041
Gray, Z., Douzandegan, Y., Tabarraei, A., & Moradi, A. (2018). Optimization of SW480 Colon Cancer Cells Transfection with Lipofectamine 2000. Medical Laboratory Journal, 12(4), 12-16.
Hameed, R. S., Waheed, M. H., & Atwan, Z. W. (2022). Upregulated CXCL10 gene Expression in SARS-CoV-2 Infected people. Basrah Journal of Science, 40(2), 357-365.
He, M., Wang, M., Huang, Y., Peng, W., Zheng, Z., Xia, N., ... & Tian, D. (2016). The ORF3 protein of genotype 1 hepatitis E virus suppresses TLR3-induced NF-κB signaling via TRADD and RIP1. Scientific reports, 6(1), 27597. https://doi.org/10.1038/srep27597
Koyama, S., Ishii, K. J., Coban, C., & Akira, S. (2008). Innate immune response to viral infection. Cytokine, 43(3), 336-341. https://doi.org/10.1016/j.cyto.2008.07.009
LeDesma, R., Nimgaonkar, I., & Ploss, A. (2019). Hepatitis E virus replication. Viruses, 11(8), 719. https://doi.org/10.3390/v11080719
Lei, Q., Li, L., Huang, W., Qin, B., & Zhang, S. (2019). HEV ORF3 downregulatesCD14 and CD64 to impair macrophages phagocytosis through inhibiting JAK/STAT pathway. Journal of Medical Virology, 91(6), 1112-1119. https://doi.org/10.1002/jmv.25400
Lhomme, S., Marion, O., Abravanel, F., Izopet, J., & Kamar, N. (2020). Clinical manifestations, pathogenesis and treatment of hepatitis E virus infections. Journal of clinical medicine, 9(2), 331. https://doi.org/10.3390/jcm9020331
Maurer, L., El Andari, J., Rapti, K., Spreyer, L., Steinmann, E., Grimm, D., & Dao Thi, V. L. (2022). Induction of hepatitis E virus anti-ORF3 antibodies from systemic administration of a muscle-specific adeno-associated virus (AAV) vector. Viruses, 14(2), 266. https://doi.org/10.3390/v14020266
Mohammed, A. J., Atwan, Z. W., & Abdul-Kareem, A. (2020). Gene Expression of Promyelocytic Leukemia Proteins and IFN-γ Is Reduced in Rotavirus-Infected Children. Iranian Biomedical Journal, 24(2), 128. https://doi.org/10.29252%2Fibj.24.2.128
Paroha, R., Chaurasiya, S. K., & Chourasia, R. (2019). Phospholipase C‐γ2 promotes intracellular survival of mycobacteria. Journal of Cellular Biochemistry, 120(4), 5062-5071. https://doi.org/10.1002/jcb.27783
Ridha, M. I., & Atwan, Z. W. (2022). Upregulation of ABL1 Gene Expression Due to the Co-expression of PML-II and adenovirus Genome Transfection. Journal of Basrah Researches (Sciences), 48(1).
Rivero‐Juarez, A., Frias, M., Martinez‐Peinado, A., Risalde, M. A., Rodriguez‐Cano, D., Camacho, A., ... & Rivero, A. (2017). Familial hepatitis E outbreak linked to wild boar meat consumption. Zoonoses and public health, 64(7), 561-565. https://doi.org/10.1111/zph.12343
Salman, A. A., Waheed, M. H., Ali Αbdulsahib, A. A., & Atwan, Z. W. (2021). Low type I interferon response in COVID 19 patients: Interferon response may be a potential treatment for COVID 19. Biomedical reports, 14(5), 1-5. https://doi.org/10.3892/br.2021.1419
Sangour, M. H., Ali, I. M., Atwan, Z. W., & Al Ali, A. A. A. L. A. (2021). Effect of Ag nanoparticles on viability of MCF-7 and Vero cell lines and gene expression of apoptotic genes. Egyptian Journal of Medical Human Genetics, 22, 1-11. https://doi.org/10.1186/s43042-020-00120-1
Sooryanarain, H., Ahmed, S. A., & Meng, X. J. (2021). Progesterone-mediated enhancement of hepatitis E virus replication in human liver cells. Mbio, 12(3), e01434-21. https://doi.org/10.1128/mBio.01434-21
Wu, C., Wu, X., & Xia, J. (2020). Hepatitis E virus infection during pregnancy. Virology Journal, 17, 1-11. https://doi.org/10.1186/s12985-020-01343-9
Wu, Z., Cheng, L., Xu, J., Li, P., Li, X., Zou, D., ... & Cao, H. (2020). The accessory protein ORF3 of porcine epidemic diarrhea virus inhibits cellular interleukin-6 and interleukin-8 productions by blocking the nuclear factor-κB p65 activation. Veterinary microbiology, 251, 108892. https://doi.org/10.1016/j.vetmic.2020.108892
Xu, R., Yu, S. S., Yao, R. R., Tang, R. C., Liang, J. W., Pang, X., & Zhang, J. (2021). Interferon-inducible LINC02605 promotes antiviral innate responses by strengthening IRF3 nuclear translocation. Frontiers in Immunology, 12, 755512. https://doi.org/10.3389/fimmu.2021.755512
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Biomedicine and Chemical Sciences
This work is licensed under a Creative Commons Attribution 4.0 International License.
