In Vitro Cytotoxic Potential and Integrated Network Pharmacology, Molecular Docking and Molecular Dynamic Approaches to Decipher the Mechanism of Gymnostachyum febrifugum Benth., in the Treatment of Breast Cancer.

Gymnostachyum febrifugum, a less-known ethnomedicinal plant from the Western Ghats of India, is used to treat various diseases and serves as an antioxidant and antibacterial herb. The present study aims to profile the cytotoxic phytochemicals in G. febrifugum roots using GC-MS/MS, in vitro confirmation of cytotoxic potential against breast cancer and an in silico study to understand the mechanism of action. Phytochemical profiling using GC-MS/MS showed the presence of eight cytotoxic molecules with lupeol in high abundance. A potent cytotoxic effect of G. febrifugum roots against breast cancer was also observed with antiproliferation, antimigration, inhibition in colony formation and death of breast cancer cells. Further, the cytotoxic potential of the plant was confirmed with the apoptosis of cells as observed in the flow cytometry. In silico network pharmacology, GO and KEGG analysis suggested the modulation of proteins of MAPK, PI3K-AKT and apoptosis pathways by lupeol to induce cytotoxicity in breast cancer. Further, dynamic simulation revealed MAPK and AKT as the major targets for lupeol. Our studies comprehensively elucidated the role of lupeol, a major phytochemical in G. febrifugum to induce cytotoxicity against breast cancer by targeting major cancer signaling pathways, providing a promising strategy and scientific basis to explore lupeol in targeted cancer therapy.