Purpose <p>Lung cancer remains the predominant cause of cancer-related mortality worldwide. Piperlongumine is a natural extract that has antitumor effects. However, the mechanism of Piperlongumine on lung cancer remains to be elucidated. This study aimed to investigate the effects and potential mechanisms of Piperlongumine on non-small cell lung cancer.</p> Methods <p>The anti-proliferative ability of Piperlongumine on lung cancer was assessed using the Cell Counting Kit-8 (CCK-8) and colony formation assays. The cell migration ability was evaluated by the wound healing assay. Additionally, western blot and immunofluorescence assays were performed to examine the expression levels of relevant proteins.</p> Results <p>Our results indicated that Piperlongumine effectively inhibited the proliferation and migration of A549 and H1299 cells. Mechanistically, Piperlongumine suppressed the phosphorylation of epidermal growth factor receptor (EGFR), focal adhesion kinase (FAK), and signal transducer and activator of transcription 3 (STAT3). It also blocked EGF-induced phosphorylation of FAK and STAT3, as well as EGF-induced cell proliferation. Furthermore, Piperlongumine reduced the expression of Cyclin D1.</p> Conclusions <p>Piperlongumine inhibits lung cancer cell proliferation and migration by modulating the EGFR/FAK/STAT3 signaling pathway. These findings suggest that Piperlongumine may serve as a potential drug for the treatment of lung cancer.</p>