Celik-Turgut, GurbetOlmez, NazmiyeKoc, TugbaOzgun-Acar, OzdenSemiz, AsliDodurga, YavuzSen, Alaattin2025-09-252025-09-2520230378-11191879-0038https://doi.org/10.1016/j.gene.2022.147099https://hdl.handle.net/20.500.12573/4590Sen, Alaattin/0000-0002-8444-376X; Ozgun Acar, Ozden/0000-0002-2910-6349; Celik Turgut, Gurbet/0000-0002-2306-6972; Satiroglu Tufan, N Lale/0000-0001-9399-0960In this study, it was aimed to elucidate the interaction between aryl hydrocarbon receptor (AHR), nuclear factor -kappa B (NF-kB), and cytochrome P4501A1 (CYP1A1) with hepatitis B virus X protein (HBX) in a human liver cancer cell line (HepG2) transfected with HBX. First, AHR, NF-kB, and CYP1A1 genes were cloned into the appropriate region of the CheckMate mammalian two-hybrid recipient plasmids using a flexi vector system. Renilla and firefly luciferases were quantified using the dual-luciferase reporter assay system to measure the interactions. Secondly, transient transfections of CYP1A1 and NF-kB (RelA) were performed into HBX-positive and HBX-negative HepG2 cells. The mRNA expression of CYP1A1 and NF-kB genes were confirmed with RT-PCR, and cell viability was measured by WST-1. Further verification was assessed by measuring the activity and protein level of CYP1A1. Additionally, CYP1A1/HBX protein-protein interactions were performed with co-immunoprecipitation, which demonstrated no interaction. These results have clearly shown that the NF-kB and AHR genes interact with HBX without involving CYP1A1 and HBX protein-protein interactions. The present study confirms that AHR and NF-kB interaction plays a role in the HBV mechanism mediated via HBX and coordinating the carcinogenic or inflammatory responses; still, the CYP1A1 gene has no effect on this interaction.eninfo:eu-repo/semantics/closedAccessHepatocellular CarcinomaHepatitis B Virus X ProteinNuclear Factor-Kappa BAryl Hydrocarbon ReceptorArticle10.1016/j.gene.2022.1470992-s2.0-85143722253