🧫 Experiment Protocol
ExploratoryTriple negative breast cancerPPARγ, GPX4, NCOA4TNBC cell linesproposed
Mechanistic studies investigating how S-equol induces ferroptosis in triple negative breast cancer cells through modulation of the PI3K/AKT/mTOR signaling pathway. The experiments examined the effects of S-equol on lipid peroxidation levels, intracellular Fe²+ concentrations, and key ferroptosis regulators including PPARγ, GPX4, and NCOA4. The study utilized specific agonists for PPARγ and STAT3, as well as autophagy inhibitor ULK101, to validate the proposed mechanism. Additionally, NCOA4 knockdown and PPARγ overexpression experiments were performed to further confirm the molecular pathways involved in S-equol-mediated ferroptosis induction.
PRIMARY OUTCOME
Ferroptosis induction markers (lipid peroxidation, Fe²+ levels)
EXPECTED OUTCOMES
Increased lipid peroxidation and Fe²+ levels, decreased GPX4 activity
SUCCESS CRITERIA
Significant changes in ferroptosis markers upon S-equol treatment
PROTOCOL
Lipid peroxidation assays, iron level measurement, gene knockdown/overexpression, pharmacological inhibition