Evaluation of deubiquiyinase USP15 expression during HCV replication

• Published in: Annals of hepatology 2022-12, Vol.27, p.100855, Article 100855
• Main Authors: Bazaldúa-Ramos, L, Rivas-Estilla, AM, Lozano-Sepúlveda, SA
• Format: Article
• Language: English
• ISSN: 1665-2681; 2659-5982
• DOI: 10.1016/j.aohep.2022.100855

This study aimed to evaluate the USP15 protein expression in HCV replication in vitro. Huh-7 cell line was transfected with pFKI plasmid that encodes for the non-structural HCV proteins; this expression is regulated by the T7 RNA polymerase promoter; therefore, prior to transfection, the cells were infected with Vaccinia Virus for T7 RNA polymerase expression. After 24 h of transfection, at 37°C and 5% CO2, total protein was extracted and quantified using the Bradford method. USP15 expression was evaluated by Western Blot assay using the antibodies for USP15, NS3-HCV and actin as a control. The expression of NS3 was found in the transfected cells; in addition, a decrease in the expression of USP15 was observed compared to the control without viral proteins. USP15 expression was shown to be downregulated in cells expressing HCV nonstructural proteins compared to control cells. A lower effect on USP15 expression was detected in the transfected cells compared to the HCV-replicon cells (positive control); this may be due to the low expression of NS3 in transfected cells. Therefore, we observed a decrease in USP15 expression dependent on NS3 expression. USP15 is known to regulate pathways including TLR signaling, RIG-I signaling, NF-kB, and IRF3/IRF7-dependent transcription to produce pro-inflammatory cytokines and type I interferons. Therefore it is important to elucidate the mechanisms involved in this regulation by HCV. USP15 expression is decreased in the presence of HCV nonstructural proteins. The resources used in this study were from the hospital without any additional financing The authors declare no potential conflicts of interest.