Introduction: Bladder cancer (Blca) is highly recurrent with limited options for advanced disease, and the pharmacological basis of traditional Chinese medicine formulas such as Xihuang Wan (XHW) in Blca is poorly defined. This study investigated the anti-bladder cancer mechanism of XHW by identifying its core bioactive component and its modulation of PI3K/AKT/BCL2-mediated apoptosis. Methods: Network pharmacology integrated TCMSP/SymMap and GeneCards/OMIM to identify active ingredients, Blca targets, and enriched pathways. Molecular docking and 100-ns molecular dynamics simulations evaluated the binding of the core component quercetin to key targets. CCK-8 and Western blot assays assessed XHW cytotoxicity, PI3K-AKT signaling, and apoptosis-related proteins in MB49 cells. Antitumor efficacy was confirmed in a C57BL/6 mouse MB49 subcutaneous xenograft model. Results: We identified 110 common targets and highlighted quercetin as a central ingredient acting on AKT1 and BCL2. KEGG analysis implicated the PI3K-AKT pathway, and docking/simulations supported stable quercetin binding. XHW inhibited MB49 proliferation and suppressed tumor growth in vivo. XHW, similar to Wortmannin, reduced phosphorylated AKT, increased the Bax/Bcl-2 ratio, and elevated cleaved caspase-3, indicating mitochondrial apoptosis via PI3K-AKT inhibition. Discussion: These findings indicate that XHW acts partly through functional inhibition of the PI3K/ AKT/BCL2 axis rather than nonspecific cytotoxicity Conclusion: Xihuang Wan exerts anti-bladder cancer activity mainly through quercetin targeting the PI3K/AKT/BCL2 axis to inhibit tumor cell survival and trigger mitochondrial apoptosis, supporting further development of XHW and quercetin as adjunctive strategies for bladder cancer.
J et al. (2026) studied this question.