Autophagy-related biological targets and network mechanisms of juglone against bladder cancer.

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.