Sanguinarine Induces ROS ‐Mediated Mitochondrial Dysfunction and Inhibits AKT / GSK3 Signaling to Potentiate Apoptotic Effects in Cutaneous T‐Cell Lymphoma

Cutaneous T-cell lymphoma (CTCL) is a rare non-Hodgkin lymphoma with limited durable treatment options and poor overall survival, underscoring the need for new therapeutic approaches. Sanguinarine (SNG), a natural benzophenanthridine alkaloid, has demonstrated anticancer activity in several malignancies, but its potential role in CTCL remains unexplored. We hypothesized that SNG induces oxidative stress and mitochondrial dysfunction, leading to the inhibition of the PI3K/AKT/GSK3 pathway and apoptosis in CTCL cells. In vitro experiments were performed using HH and H9 CTCL cell lines. Cytotoxicity, apoptosis, and mitochondrial function were evaluated by cell viability assays, caspase activation, PARP cleavage, Bax/Bcl-2 ratio, mitochondrial membrane depolarization, and cytochrome c release. ROS generation and glutathione depletion were measured with and without N-acetyl cysteine (NAC) rescue. Western blotting assessed modulation of the AKT/GSK3α/β/mTOR pathway and expression of anti-apoptotic proteins (XIAP, cIAPs, Mcl-1). Synergistic effects with bortezomib (BTZ) were analyzed. Network pharmacology, molecular docking, molecular dynamics (MD) simulations, and binding free energy (BFE) calculations were used to identify SNG's molecular targets. SNG reduced CTCL cell viability (IC₅₀ < 5 μM) and triggered mitochondrial-mediated apoptosis, accompanied by Bax/Bcl-2 modulation, ΔΨm loss, cytochrome c release, and caspase-9/3 activation. ROS accumulation and glutathione depletion contributed to cytotoxicity, effects that were reversed by NAC. SNG suppressed AKT/GSK3/mTOR signaling and downregulated anti-apoptotic proteins. Notably, SNG enhanced the anticancer activity of BTZ in combination studies. Computational analyses supported AKT and Bcl-2 as key binding targets. SNG exerts potent anticancer effects in CTCL by inducing ROS-dependent mitochondrial apoptosis and inhibiting the PI3K/AKT/GSK3 signaling pathway. Its synergy with BTZ and computational validation of AKT/Bcl-2 targeting underscore its potential as a novel therapeutic candidate for CTCL, warranting further preclinical investigation.