Pharmacological Inhibition of APT2 by Solasonine Promotes Ferroptosis Through Palmitoylation Modulation in Gallbladder Cancer.

Gallbladder cancer (GBC), the most prevalent and lethal biliary tract malignancy, lacks effective therapeutic strategies. Solasonine (SS), a bioactive steroidal alkaloid derived from Solanum nigrum L, exhibits antitumor activity in multiple cancers, but its role in GBC and association with ferroptosis remains unknown. The purpose of this study is to investigate the therapeutic effects of SS on GBC and its underlying mechanisms using both in vitro and in vivo models. The efficacy was tested in subcutaneous models. Then, the anti-GBC effects of SS were assessed via CCK-8, colony formation, and EdU assays. Biosafety was evaluated using L-2F7 cells and murine serum biomarkers. RNA sequencing identified key pathways, validated by ChIP-qPCR and dual-luciferase reporter assays. Ferroptosis induction was confirmed by measuring MDA, GSH, ROS, Fe2+, qRT-PCR/Western blot (GPX4/SLC7A11), and rescue experiments. Direct binding of SS to APT2 was demonstrated through molecular docking, CETSA, and SPR. The study employed orthotopic xenograft models, transwell assays, and wound healing assays to investigate the effects of SS on the metastasis of GBC. SS triggered ferroptosis in GBC by elevating MDA, ROS, and Fe2+ while depleting GSH and downregulating GPX4/SLC7A11. Mechanistically, SS directly bound APT2, inhibiting its enzymatic activity and blocking STAT3 depalmitoylation. This prevented p-STAT3 nuclear translocation, suppressing transcription of ferroptosis inhibitors GPX4 and SLC7A11. In vivo, SS suppressed tumor growth by subcutaneous and orthotopic models with no systemic toxicity. Our findings reveal SS as a novel APT2-targeted ferroptosis inducer that inhibits GBC progression via inhibiting the depalmitoylation process of STAT3, providing a safe and effective therapeutic candidate for this intractable malignancy.