Network pharmacology-guided identification of Rabdosia rubescent as a suppressor of laryngeal cancer via MMP9-mediated extracellular matrix remodeling: integrated in silico and experimental validation
Background Laryngeal cancer, particularly laryngeal squamous cell carcinoma (LSCC), poses a significant threat to global health, especially within Asian populations. While Rabdosia Rubescens is known for its antitumor properties, the specific bioactive compounds and mechanisms underlying its activity against laryngeal carcinoma remain to be fully elucidated. Methods This study employed an integrated approach combining network pharmacology, molecular docking, and 500 ns molecular dynamics simulations to screen bioactive compounds from Rabdosia Rubescens. Key therapeutic targets were identified and validated using transcriptomic data from The Cancer Genome Atlas (TCGA). The most promising candidate was subsequently validated in vitro using cytotoxicity assays (TU177 and RK33 cell lines) and wound healing migration assays, alongside MMP9 knockdown models to confirm mechanistic specificity. Results Network pharmacology identified MMP9 as a critical therapeutic target, which was found to be significantly overexpressed in LSCC tissues. Virtual screening and MD simulations identified Cirsiliol as the most potent inhibitor among the tested phytochemicals, exhibiting a highly favorable binding free energy (ΔGbinding = −179.793 kJ/mol) and stable interactions with key residues (ALA189, HIS266, and LEU243). Experimental validation demonstrated that Cirsiliol induced dose-dependent growth inhibition with IC50 values of 31.59 μM in TU177 cells and 17.73 μM in RK33 cells. Furthermore, the compound significantly suppressed cell migration, mirroring the effects observed in MMP9-knockdown TU177 cells. Conclusion This study establishes Cirsiliol as a potent, specific inhibitor of MMP9 derived from Rabdosia Rubescens. By effectively targeting the MMP9 pathway, Cirsiliol suppresses tumor invasiveness and migration, highlighting its potential as a promising lead compound for the treatment of laryngeal cancer.