Abstract: This study aimed at investigating the effects and mechanisms of 7-O-succinyl macrolactin A in inhibiting human lung cancer. After treatment of human lung cancer cell lines H460 with 7-O-succinyl macrolactin A, MTS assay was employed to determine cell proliferation; crystal violet staining was used to detect cell adhesion of H460; transwell chamber assay and wound healing assay were performed to evaluate cell invasion and migration; and flow cytometry assay was adopted to evaluate cell cycle. Western blotting and real-time PCR were also employed to determine the expression of β-catenin, c-Myc, Cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2, Survivin and MMP-2/9. The phosphorylation of AKT and mTOR was determined as well. In vitro proliferation of H460 was inhibited significantly by 7-O-succinyl macrolactin A. Cell adhesion, invasion and migration abilities were also attenuated. Western blot and real-time PCR showed that the expressions of β-catenin, c-Myc, cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2, Survivin and MMP-2/9 were down-regulated by 7-O-succinyl macrolactin A. It was also found that phosphorylation of AKT and mTOR was inhibited by 7-O-succinyl macrolactin A. 7-O-succinyl macrolactin A can inhibit the in vitro growth and invasion of human lung cancer cell lines H460.