Role of AHR, NF-kB and CYP1A1 crosstalk with the X protein of Hepatitis B virus in hepatocellular carcinoma cells

• Published in: Gene Vol. 853; p. 147099
• Main Authors: Celik-Turgut, Gurbet, Olmez, Nazmiye, Koc, Tugba, Ozgun-Acar, Ozden, Semiz, Asli, Dodurga, Yavuz, Lale Satiroglu-Tufan, Naciye, Sen, Alaattin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.02.2023
• Subjects: Animals; Aryl hydrocarbon receptor; Carcinoma, Hepatocellular - pathology; Cell Line; Cytochrome P-450 CYP1A1 - genetics; Cytochrome P4501A1; Hepatitis B virus - metabolism; Hepatitis B virus X protein; Hepatocellular carcinoma; Humans; Liver Neoplasms - pathology; Mammals - metabolism; NF-kappa B - metabolism; Nuclear factor-kappa B; Receptors, Aryl Hydrocarbon - genetics; Receptors, Aryl Hydrocarbon - metabolism; Signal Transduction; Nuclear factor-kappa B; PAS; DTT; HLH; NF-kB; TCDD; Hepatocellular carcinoma; 3MC; AP-1; NFAT; BSA; SD; IFN-γ; CREB; TBK1; ε-ACA; FBS; RT-PCR; HepG2; HBV; Cytochrome P4501A1; HBX; BCA; XREs; EROD; c-Myc; STAT3; HCC; AhR; Co-IP; EDTA; WST-1; IL-6; PMFS; EMEM; Aryl hydrocarbon receptor; SDS-PAGE; ATF; MEM; Hepatitis B virus X protein; B(a)P; ROS; CYP1A1; GAPDH
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2022.147099

[Display omitted] •HBX overexpression in HepG2 cells leads to activation of NF-kB and AhR signaling pathways.•Depending on the HBX-AhR interaction, HBX-mediated hepatocarcinoma may develop in liver cells.•HBX may coordinate direct interactions AhR, and NF-kB, and indirect interactions with CYP1A1. In 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.