Icariside II induces ferroptosis and apoptosis by activating oxidative stress for non-small-cell lung cancer therapy.

Treatment strategies focusing on the induction of multiple programmed cell death (PCD) have gained attention in non-small-cell lung cancer (NSCLC). PCD represented by apoptosis and ferroptosis is closely linked to oxidative stress. This study aimed to investigate whether icariside II (ICS II) induces dual PCD and elucidate the underlying mechanisms. The current study showed that ICS II inhibited cell progression and metastasis and was more selective towards NSCLC cells. Transcriptomic analysis revealed that ICS II induced ferroptosis, characterized by excessive levels of reactive oxygen species (ROS), depletion of reductive compounds, and mitochondrial impairment. Functional analysis revealed that ICS II suppressed Nrf2-mediated SLC7A11/GPX4/HO-1 transcription and activated ACSL4-mediated lipid peroxidation, which were rescued by ferrostatin-1 and Nrf2 overexpression. Simultaneously, ICS II activated caspase family proteins and reduced PARP activity, thereby triggering apoptosis. N-acetyl-L-cysteine abolished the effects of ICS II on NSCLC cells and restored cell viability, confirming the functional contribution of ROS in ICS II-induced PCD. Finally, a mouse subcutaneous tumor model was established to assess the drug efficacy and toxicity. As expected, ICS II inhibited tumor development in a mouse subcutaneous tumor model with minimal toxicity. Taken together, ICS II exerts anti-NSCLC efficacy by weakening cellular antioxidative capacity (inhibition of Nrf2 activity) and promoting the accumulation of substrates for lipid peroxidation (ACSL4 activation), triggering oxidative stress, and exhibiting dual PCD induction. Our findings demonstrate the potential of ICS II for use in clinical NSCLC therapy.