Objective:To investigate the effect and mechanism of longikaurin A (LK-A) on glioma proliferation.Methods:Resuscitated frozen human glioma cell lines U87 and SNB19 were in vitro sub-cultured; CCK-8 assay was used to detect the cell activity changes 24 and 48 h after 0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 6.0, 8.0 μmol/L LK-A. Edu proliferation assay was used to detect the changes in number of Edu positive cells after 0, 1, 2 and 3 μmol/L LK-A, and cell clonal formation assay was used to detect the changes in number of cell colony formation after 0, 0.1, 0.2 and 0.3 μmol/L LK-A. Flow cytometry was used to detect the changes in cell cycle proportion after 0, 1, 2 and 3 μmol/L LK-A. Expression changes of proliferation-related proteins (Ki-67 and c-Myc) and cell cycle-related proteins (Cyclin B1, Cyclin D1, cyclin dependent kinase 1 [CDKl]) and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway-related proteins were detected by Western blotting after 0, 1, 2, and 3 μmol/L LK-A. Results:U87 and SNB19 cell viabilities decreased gradually after being treated with 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 6.0, and 8.0 μmol/L LK-A for 24 and 48 h compared with those with 0 μmol/L LK-A, with significant differences ( P<0.05); cell viability was dose-dependent. The number of Edu positive cells in U87 and SNB19 after being treated with 2, and 3 μmol/L LK-A was significantly decreased compared with that with 0 μmol/L LK-A ( P<0.05). The colony formation number of U87 and SNB19 cells after being treated with 0.1, 0.2 and 0.3 μmol/L LK-A decreased significantly compared with that with 0 μmol/L LK-A ( P<0.05). The proportion of U87 and SNB19 cells at G2/M phase after being treated with 2 and 3 μmol/L LK-A were increased significantly compared with that with 0 μmol/L LK-A ( P<0.05). The Ki-67, c-Myc, Cyclin B1, CDK1, p-PI3K and p-AKT expressions were significantly decreased, while Cyclin D1 expression was significantly increased in U87 and SNB19 cells after being treated with 2 and 3 μmol/L LK-A compared with those with 0 μmol/L LK-A ( P<0.05). Conclusion:LK-A can inhibit the glioma proliferation and arrest glioma at G2/M phase, with dose-dependent manner; the mechanism is related to inhibition of PI3K/AKT signaling pathway by LK-A.