Glycycoumarin Induces G2/M Cell Cycle Arrest and Apoptosis in Non-Small-Cell Lung Cancer Cells via Suppressing the PI3K/AKT/mTOR/HIF-1α Pathway.

Current therapies for Non-Small Cell Lung Cancer (NSCLC) are frequently compromised by drug resistance and adverse effects. Glycycoumarin (GCM), a natural coumarin derived from licorice, has been shown to exhibit anticancer properties. Nevertheless, its mechanisms of action against NSCLC remain ambiguous. This study aimed to investigate the antitumor effects of GCM on NSCLC and clarify the underlying molecular mechanism through an integrative approach. The bioactivity of GCM on NSCLC cells was assessed using crystal violet staining, colony formation, migration, and invasion assays, as well as flow cytometry. In vivo efficacy was evaluated utilizing xenograft tumor models. Network pharmacology, molecular docking, and dynamic simulation were employed for mechanistic prediction, followed by experimental validation through Western blot and RT-qPCR. GCM induced G2/M phase arrest, apoptosis, and a significant inhibition of A549 cell proliferation. Subsequent in vivo investigations demonstrated that GCM administration suppressed the growth of transplanted tumors in the A549 xenograft model. Network pharmacology analysis revealed that PI3K/AKT and HIF-1 signaling were important mediators, with AKT1, mTOR, PIK3CA, Bcl2, and HIF-1α among the core targets. The stable binding of GCM to these targets was predicted using molecular docking and dynamics. Experimental validation showed that GCM inhibited the progression of NSCLC by controlling the expression of the PI3K/AKT/mTOR/HIF-1α pathway. This is the first study to demonstrate the anticancer efficacy of GCM in NSCLC. GCM appears to act on multiple targets within the PI3K/AKT/mTOR/HIF-1α axis, which may offer advantages in overcoming therapeutic resistance. The multi-target mechanism of action, combined with its safety profile, supports GCM as a potential agent for further preclinical evaluation in NSCLC. GCM impedes NSCLC progression by inducing cell cycle arrest and apoptosis, potentially via the suppression of the PI3K/AKT/mTOR/HIF-1α signaling axis.