An increasing number of studies have demonstrated that phillygenin (PG) exerts anti-oxidant, anti-inflammatory and anti-cancer activities. However, the effects of PG on the proliferation and invasion in non-small cell lung cancer (NSCLC) cells have not been clarified. In this study, MTT assay and flow cytometry were conducted to investigate the effect of PG on proliferation and apoptosis of NSCLC cells in vitro, respectively. A xenograft model of A549 cell was established in nude mice to validate the in vitro findings. Western blot were performed to measure the expression of molecules involved in AMPK/ERK/NF-κB pathway. Results suggested that PG (50 or 100 μM) was significantly cytotoxic to A549 cells and SPC-A1 cells in vitro. PG treatment also inhibited the tumor growth of NSCLC cell mouse xenografts in vivo. These anti-proliferative and pro-apoptosis effects of PG were found to be regulated by the AMPK/ERK/NF-κB pathway. Consequently, PG suppressed proliferation and induced cell apoptosis in NSCLC cells. In conclusions, PG regulates AMPK/ERK/NF-κB axis in NSCLC cells, thereby inhibiting the proliferation and promoting the apoptosis of NSCLC cells.