Abstract
OBJECTIVE: This study aimed to explore juglone's mechanism in inducing autophagy and apoptosis in bladder cancer (BLCA) via network pharmacology and experimental validation. METHODS: Juglone's effects on BLCA cell proliferation, apoptosis, and autophagy were assessed using CCK-8, flow cytometry, transmission electron microscopy, and Western blotting. Network pharmacology, molecular docking, and dynamics simulations identified key targets. In vivo validation employed H&E, immunohistochemical, and TUNEL staining. RESULTS: Juglone suppressed T24 and UMUC-3 cell proliferation, enhanced autophagy markers, and induced apoptosis. Network analysis identified 108 shared targets, with AKT1, CASP3, and TP53 as core nodes. Pathway enrichment implicated the PI3K/Akt signalling pathway, supported by molecular docking. Autophagy inhibition reduced juglone-induced apoptosis, confirming autophagic death's role. CONCLUSIONS: Juglone triggers BLCA autophagy via PI3K/AKT/mTOR, upregulates apoptotic proteins, and activates caspase 3, promoting apoptosis.