AIM: To investigate the effects of isonlosinine on proliferation, invasion, migration and autophagy of PC9 cells in non-small cell lung cancer (NSCLC), and to explore its possible molecular mechanism. METHODS: The effect of Isoliensinine on the proliferation of PC9 cells were measured by CCK-8 assay, and the IC50 value of PC9 cells was calculated. Wound healing and transwell experiments were used to study the effect of Isoliensinine on migration and invasion of PC9 cells in vitro, respectively. The formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The expression levels of LC3, p-ERK and ERK in the PC9 cells were determined by western blot. RESULTS: Isonlosinine significantly inhibited the proliferation of PC9 cells. IC50 of isonlosinine (24 h) for the PC9 cells was 34.11 μmol/L. Isonlosinine significantly inhibited cell migration and invasion of PC9 cells. The results of acridine orange fluorescent staining showed that the number of the intracellular acid dye follicular bright red fluorescence in PC9 cells was significantly increased after isonlosinine treatment, while the autophagic lysosomes were rarely observed in control group. The expression of LC3-II in PC9 cells was significantly enhanced after isonlosinine treatment. Furthermore, molecular mechanism study showed that isonlosinine could activate the expression level of p-ERK. CONCLUSION: Isoliensinine significantly inhibits the proliferation, migration and invasion, and induces autophagy of PC9 cells, which may be correlated with the activation of ERK signaling pathway.