Dual inhibition of MMP-2 and actin dynamics by a novel bis-chalcone: an anticancer strategy for oral squamous cell...

Oral squamous cell carcinoma (OSCC), accounts for approximately 90% of oral cancer cases, and due to its aggressive nature and limited treatment options remains a significant clinical challenge. In the pursuit of novel therapeutic agents, bis-chalcones (a subclass of chalcones) have emerged as promising candidates with multipronged anticancer potential. In this study, we synthesized and evaluated a novel bis-chalcone (B2OCH₃), for its cytotoxic, anti-migratory, anti-invasive, and anti-adhesive activities, as well as potential mechanisms of action. The compound was obtained via Claisen-Schmidt condensation and spectroscopically characterized. Cytotoxicity was assessed using MTT, pulse treatment, and trypan blue exclusion assays. Morphological changes were examined with panoptic staining and immunofluorescence. Clonogenic, migration, adhesion, and invasion assays were performed to assess anti-metastatic potential. Molecular docking and gene/protein expression analyses were conducted by quantitative PCR and Western blot. B2OCH₃ exhibited potent cytotoxicity against HSC-3, SCC-4, and CAL-27 OSCC cell lines, with HSC-3 being the most sensitive. The compound demonstrated selectivity for cancer cells as compared with non-tumor HS5 and HaCaT cells. Reduced viability was accompanied by mitotic figures and apoptotic bodies. B2OCH₃ significantly inhibited colony formation, migration, invasion, and adhesion in HSC-3 cells. Morphological analysis revealed modulation of actin filament organization and reduced filopodia formation. Molecular docking indicated MMP-2 and F-actin affinity, and was corroborated by molecular assays confirming protein modulation for both. Our findings identify B2OCH₃ as a promising therapeutic candidate for OSCC, highlighting its potential to simultaneously target both MMP-2 activity and actin cytoskeleton regulation.