Paris saponin Ⅶ (PSⅦ),a bioactive constituent extracted from Trillium tschonoskii Maxim.,is cytotoxic to several cancer types.This studywas designed to explore whether PSⅦ prevents non-small-cell lung cancer (NSCLC) proliferation and to investigate its molecular target.AMP-activated protein kinase (AMPK) has been implicated in the activation of autophagy in distinct tissues.In cultured human NSCLC cell lines,PSⅦ induces autophagy by activating AMPK and inhibiting mTOR signaling.Furthermore,PSⅦ-induced autophagy activation was reversed by the AMPK inhibitor compound C.Computational docking analysis showed that PSⅦ directly interacted with the allosteric drug and metabolite site of AMPK to stabilize its activation.Microscale thermophoresis assay and drug affinity responsive target stability assay further confirmed the high affinity between PSⅦ and AMPK.In summary,PSⅦ acts as a direct AMPK activator to induce cell autophagy,which inhibits the growth of NSCLC cells.In the future,PSⅦ therapy should be applied to treat patients with NSCLC.