Ginsenoside F2 inhibits MLCK to induce synthetic lethality in MYC-driven triple-negative breast cancer and pancreatic cancer

Myosin Light Chain Kinase (MLCK) represents a synthetic lethal interaction with the “undruggable” oncoprotein MYC. In this study, we identified Ginsenoside F2 (GF2) as a novel MLCK inhibitor through molecular docking-based virtual screening of a natural compound library, followed by in vitro cellular validation. GF2 treatment inhibited MLCK kinase activity, as demonstrated by the reduced phosphorylation of its substrate, myosin II regulatory light chain (MLC). The direct binding of GF2 to MLCK was confirmed using the cellular thermal shift assay (CETSA), which revealed decreased MLCK thermotolerance after GF2 treatment. Notably, GF2 selectively induced apoptosis in MYC-transformed cells while sparing normal counterparts. Triple-negative breast cancer (TNBC) and pancreatic cancer cells with high MYC expression are sensitive to GF2 treatment. Moreover, combining GF2 with the Bcl2 inhibitor venetoclax synergistically enhanced apoptosis in MYC-driven cancer cells. These findings establish GF2 as a novel MLCK inhibitor and underscore the therapeutic potential of targeting MLCK in MYC-driven malignancies, particularly TNBC and pancreatic cancer.