Trigonelline suppresses the tumor progression via STAT2 signalling pathway in non-Small cell lung carcinoma

Trigonelline, a naturally derived bioactive compound, has gained interest for its potential anticancer properties. This study investigates its therapeutic efficacy against A549 lung cancer cells and elucidates the underlying molecular mechanisms. Cytotoxic effects were evaluated using MTT and Trypan blue exclusion assays in A549 cancer cells and biocompatibility were examined in 3T3-L1 mouse fibroblast cells. Flow cytometry was employed to analyze cell cycle distribution, apoptotic induction, and metabolic alterations were utilised using specific metabolic assay kits. Network analysis through the STRING database was performed to identify key signaling pathways affected by trigonelline. Gene expression changes were validated using RT-PCR, while Western blotting assessed protein levels of STAT2, BCL2, and BAX. Docking studies using AutoDock examined the binding affinity of trigonelline with STAT2-related targets, and molecular dynamics simulations evaluated the stability of trigonelline-bound STAT2 and BCL2 complexes using GROMACS package. Trigonelline exhibited significant cytotoxicity in A549 cells while showing minimal cell inhibitory effects in 3T3-L1 cells, indicating selective anticancer activity. Flow cytometry revealed that trigonelline induced G0/G1 cell cycle arrest and promoted apoptosis. Network and pathway analyses highlighted STAT2-mediated signaling, apoptosis, and metabolic regulation as major pathways influenced by trigonelline. RT-PCR results showed downregulation of genes involved in glycolysis and apoptosis. Western blot analysis confirmed decreased expression of STAT2 and the anti-apoptotic protein BCL2, alongside increased BAX protein levels. Docking studies demonstrated strong interactions with STAT2-associated targets, and molecular dynamics simulations confirmed the stable binding of trigonelline to STAT2 and BCL2. Trigonelline effectively inhibits the proliferation of A549 lung cancer cells by inducing cell cycle arrest, triggering apoptosis, modulating metabolic, and STAT2-mediated pathways. These findings suggest that trigonelline holds promise as a potential therapeutic candidate for lung cancer. Further in vivo studies and clinical investigations are warranted to explore its therapeutic potential.