Antiproliferative and proapoptotic effects of Dalbergia odorifera flavonoids in cSCC

Cutaneous squamous cell carcinoma (cSCC) lacks effective and well-tolerated pharmacological options for long-term management, highlighting the need for multi-target natural agents. In this study, we hypothesized that a flavonoid-rich extract from Dalbergia odorifera exerts (DOE) anti-cSCC effects by regulating apoptosis-related pathways. To test this hypothesis, we employed an integrated strategy combining UPLC-based chemical profiling, network pharmacology prediction, and in vitro and in vivo validation. UPLC analysis identified five major flavonoids in DOE-Luteolin, Naringenin, Butein, Liquiritigenin, and Formononetin. Network pharmacology predicted the involvement of key pathways, including PI3K/AKT, MAPK, and EGFR. Experimental validation, however, was focused on apoptosis-related markers Bax and Bcl-2, providing partial support for the predicted mechanisms. In vitro, DOE significantly reduced cell viability and colony-forming ability. Migration was decreased by more than 80%, and apoptosis increased substantially at higher doses. These findings were confirmed by qRT-PCR and Western blot analyses, which showed a downregulation of Bcl-2 and upregulation of Bax, consistent with the proapoptotic mechanism predicted by network pharmacology. In vivo, using a DMBA/croton oil-induced cSCC mouse model, DOE administration resulted in dose-dependent tumor inhibition and improved body weight loss, thymus, and spleen indices. DOE, as a flavonoid extract, significantly inhibits cell proliferation, migration, and promotes apoptosis in cSCC, positioning it as a promising candidate for further development as a complementary or adjuvant therapeutic strategy.