Abstract Background Nuclear factor erythroid 2– related factor 2 (NRF2) is a transcription factor and master regulator in multiple metabolic as well as physiological process. It plays a significant role in cancer progression and chemoresistance development. It induces the expression of a large number of cytoprotective proteins with antioxidant and detoxifying roles. NRF2 inhibition by ROS generating biomolecule, Alantolactone (ALT) can be potential approach for ROS-mediated cancer cell death. Methods To study this, we performed in silico and in vitro studies. In in silico studies, we used predicted NRF2 structure by AlphaFold, performed its molecular docking, and molecular dynamic (MD) simulations with ALT. Whereas, in in vitro studies, we performed western blotting to check its translocation into nucleus and qPCR to check its transcriptional activity. We performed Frontier molecular orbitals (FMOs) analysis as well to check reactivity and bioactivity of ALT. Results Molecular docking results revealed that ALT strongly binds to DNA binding domain of NRF2 with stable molecular dynamics. In vitro studies showed a significant reduction in nuclear NRF2 in taxol resistant cell line (HEY-T30) treated with ALT (30 µM) for 24 h. Our qPCR analysis showed that ALT failed to modulate mRNA expression of NRF2 downstream target genes including glutathione peroxidase-4 (GPX-4) and glutathione reductase (GSR) in MCF-7 cells. Moreover, ProTox-II profiling showed that ALT is non-toxic to other organs and can be safe if it is swallowed. FMOs method showed the reactivity and bioactivity of ALT. Conclusion ALT can significantly inhibit the NRF2 activity. The inhibition of NRF2 translocation into nucleus might help to reduce the antioxidant enzymes synthesis leading to ROS mediated cell death. Further in vitro and in vivo studies are required to investigate the underlying mechanism in-depth.
Gul et al. (Wed,) studied this question.