Phytochemicals, withaferin A and carnosol, overcome pancreatic cancer stem cells as c-Met inhibitors.

Receptor tyrosine kinases (RTKs) are pharmaceutically attractive targets due to their fundamental role in tumor formation. The hallmark of pancreatic cancer is its high mortality rate attributed to the existence of cancer stem cell (CSC) subpopulations which result in therapy resistance and recurrence. c-Met is a known pancreatic CSC marker that belongs to the family of RTKs. To surmount the hurdles related to ligand-independent c-Met activation, we aimed to elucidate the inhibitory mechanisms of withaferin A (WA) and carnosol (CA) as two hit phytochemicals against c-Met kinase domain. Both tested compounds attenuated HGF-mediated proliferation across various established c-Met+ cancer cell lines and altered cell cycle distribution accompanied by apoptosis induction. Scratch assay confirmed the anti-migratory activity of WA and CA in AsPC-1 cells. The blockade of HGF-driven cellular growth and motility was reflected by the suppression of c-Met phosphorylation and its downstream pro-survival pathway Akt. Further studies showed that the administration of WA and CA diminished the sphere-formation and clonogenic potential which was validated by down-regulation of pluripotency maintaining genes (oct-4 and nanog), demonstrating their potentiality to target pancreatic CSCs. As more than 60% of anti-cancer drugs are composed of natural product-derived inhibitors known as fourth generation inhibitors, our present data suggest that WA and CA may hold promise to eradicate CSCs in c-Met-dependent cancers.