Lung cancer, as a highly malignant neoplasm, leads to elevated morbidity and mortality rates owing to the absence of reliable approaches for prompt diagnosis and intervention. It is crucial to investigate the mechanisms underlying lung cancer development and identify biomarkers to achieve early detection and treatment. In this study, DEP domain containing 1 (DEPDC1), was identified as a hub gene with higher expression in lung adenocarcinoma (LUAD) through weighted gene co-expression network and protein-protein interaction analyses. Patients with high DEPDC1 expression had worse survival rates, and gene set enrichment analysis revealed that DEPDC1 may participate in oncogenic progression via NF-κB-mediated signaling pathways, including the cell cycle and epithelial-mesenchymal transition. DEPDC1 knockdown significantly inhibited cancer cell proliferation and motility. Further, we demonstrated that an ionizable lipid nanoparticle delivering DEPDC1 small interfering RNA effectively inhibited tumor growth in vivo, with few systemic adverse effects. Collectively, our current investigation illustrates that DEPDC1 is a potential target for LUAD therapy.