Abstract
Abstract Karanjin is a furanoflavanoid that demonstrates selective cytotoxic effects toward various cancer cells; however, its underlying mechanism has yet to be thoroughly investigated. This study aimed to elucidate the molecular mechanisms underlying karanjin-induced cytotoxicity across cancer (HeLa, MCF-7, A549, and HCT116) and healthy (HaCaT) cell lines. A combined approach integrating network pharmacology, molecular docking, and in vitro functional analyses was employed. The in vitro data showed that a high accumulation of intracellular reactive oxygen species (ROS) induced oxidative-stress-related toxicity events and subsequent cell death in cancer cells. The mitochondrial membrane potential (MMP), ATP levels, cytochrome C, and caspase 3 were more significantly affected in cancer cells (MCF-7, A549, and HCT116) as compared to healthy cells (HaCaT). Interestingly, HeLa cells were not responsive to karanjin treatment. Correlation and regression analyses indicated that mitochondrial depolarization was strongly associated with apoptosis, suggesting that mitochondrial dysfunction plays a key role downstream of oxidative stress. Network pharmacology analysis showed significant enrichment in PI3K-Akt signalling pathway, ROS regulation, and various cancer-related pathways. This suggests that ROS and the PI3K-AKT-mTOR pathway could be targets of karanjin in cancer therapy, which is supported by molecular docking and in vitro data. In conclusion, karanjin induced ROS accumulation and mitochondrial dysfunction in susceptible cancer cells (MCF-7, A549, and HCT116) in a dose-dependent manner without affecting healthy cells.