Ferroptosis-targeting compounds modulate cancer cell cytotoxicity and migration: Insights from in vitro and in silico...

Ferroptosis is an iron-dependent, non-apoptotic form of metabolic cell death driven by lipid peroxidation. While Glutathione Peroxidase 4 (GPX4) is the canonical ferroptosis suppressor, recent evidence highlights additional selenoproteins, including Thioredoxin Reductase 1 (TXNRD1) and Peroxiredoxin 6 (PRDX6), as critical modulators of ferroptotic sensitivity. We compared three mechanistically distinct ferroptosis inducers, RSL3 (a covalent GPX4 inhibitor), auranofin (a TXNRD1 inhibitor), and artesunate (ART; a pro-oxidant derivative of artemisinin), in MDA-MB231, A549 and HepG2 cell lines.

Cell viability (MTT) and wound-healing assays quantified cytotoxic and anti-migratory effects. Our inhibitor study using Ferrostatin-1(Ferro) and Liproxstatin-1(Lipo), in combination with ferroptosis inducers, confirmed the specificity of ferroptosis. Beyond GPX4, TXNRD1 and PRDX6 constitute a complementary selenium-dependent axis safeguarding cancer cells from ferroptosis. Dual targeting of GPX4 and TXNRD1, or disruption of PRDX6-mediated selenium trafficking, potentiates ferroptosis death and impedes metastatic traits.

Using in silico methods, we confirmed the interaction between drug and protein molecules. Among the tested compounds, RSL3 and auranofin exhibited strong binding affinity towards all the targeted proteins, including GPX4, TXNRD1, and PRDX6, suggesting their potential as effective ferroptosis pathway inhibitors.

These findings nominate multi-selenoproteins inhibition as a promising strategy to overcome ferroptosis resistance.