Hepatocellular carcinoma (HCC) is a common cancer worldwide with high morbidity and mortality. Ginkgolic acid (GA) is a natural compound obtained from leaves and seed coats of Ginkgo biloba L., and it has been reported to have various bioactivities. However, the effects of GA on HCC cell cycle distribution and the mechanisms involved are still unknown. By CCK-8 assay and Transwell assay, the cell viability and migration of HCC cells were shown to be inhibited significantly by GA in a concentration-dependent manner. By cell cycle analysis and western blot, the cell cycle arrest at G0/G1 phase was shown to contribute to the inhibitory effects of GA. Furthermore, the phosphorylation of p38 MAPK was found to be elevated upon GA treatment as analyzed by western blot. Thus, GA inhibited cell proliferation and migration of HCC cells by inducing G0/G1 cell cycle arrest via p38 MAPK activation, indicating GA as an agent candidate for HCC treatment.