The influence of TNF-a on the expression profile of key enzymes of steroidogenesis in H295R cells

Introduction: Tumor necrosis factor-a (TNF-a) plays an extremely important role in the regulation of hypothalamicpituitary-adrenal axis. It is believed that chronic inflammation is the main cause of cancerogenesis and TNF-a plays a significant role in both of these processes. Unfortunately, the function of TNF-a in human adrenal steroidogenesis has not been explained enough. Aim: To evaluate the changes in transcriptional activity of STAR, CYP11A1, CYP11B1, and CYP11B2 in H295R cell line exposed to TNF-a. Material and methods: NCI-H295R, human adrenocortical cell line was exposed to human recombinant TNF-a at the concentrations ranging from 0.001 to 10 nM for 3, 12, 24, and 48 h. Cells not exposed to TNF-a were the control of this experiment. RTqPCR assay was used to determine the changes in the expression of genes encoding STAR, CYP11A1, CYP11B1, and CYP11B2. Results: The highest differences between stimulated and non-stimulated cells were observed in the expression of STAR (FC = +2.2; 0.01 nM of TNF-a; 48 h); CYP11A1 (FC = +3.5; 0.1 nM of TNF-a; 24 h); CYP11B1 (FC = +7.0; 10 nM of TNF-a; 48 h); CYP11B2 (FC = +2.5; 10 nM of TNF-a; 48 h). Statistically significant differences (p < 0.05) in the expression were found only for CYP11A1. The interaction effect between genes was also noticed (p < 0.05). Conclusions: The research showed the impact of TNF-a on the expression of the key genes encoding enzymes involved in adrenal steroidogenesis. Different expression patterns of was observed, depending on time and TNF-a concentration increased synthesis of this pro-inflammatory cytokine may intensify adrenal steroidogenesis..