Background <p>Dauriporphine is a major ingredient of <i>Manispernum daericum</i> DC., which has been demonstrated to show wide anti-tumor activities. miR-424-5p, as a regulator of lung cancer, was hypothesized to serve as the therapeutic target for dauriporphine This study evaluated the potential of dauriporphine in treating lung adenocarcinoma and revealed the underlying molecular mechanism.</p> Results <p>The anti-tumor effect of dauriporphine on lung adenocarcinoma was assessed in A549 cells, and it was found that dauriporphine significantly inhibited the viability of A549 cells in a concentration-dependent manner with the half maximal inhibitory concentration (IC₅₀) value of 10.57 µM. Dauriporphine induced decreasing cell growth, motility, and energy metabolism, indicating the anti-tumor effect of dauriporphine on A549 cells. Dauriporphine inducing elevated miR-424-5p levels, while silencing miR-424-5p significantly recovered cell viability, migration, and energy metabolism of A549 cells. Mitogen-activated protein Kinase 14 (MAPK14) was negatively regulated by miR-424-5p, and the knockdown of MAPK14 could reverse the protective effect of miR-424-5p on dauriporphine-treated A549 cells.</p> Conclusion <p>Dauriporphine inhibited cell growth, metastasis, and glycolysis-related energy metabolism of lung adenocarcinoma cells via modulating miR-424-5p/MAPK14 axis. Dauriporphine can be considered in drug development for lung adenocarcinoma.</p> Clinical trial number <p>Not applicable.</p>