To investigate the effects of ginkgolide B on the proliferation and apoptosis of esophageal cancer cells and the related mechanisms. Human esophageal cancer OE19 cells were cultured in vitro. They were divided into control group (no intervention), low/medium/high dose test group (adding 6.25, 12.50 and 25.00 μmol/L ginkgolide B, respectively), ginkgolide B group ( 12.50 μmol/L ginkgolide B), positive drug group (4 mg/L cisplatin) and inhibitor group［12.50 μmol/L ginkgolide B+10.00 μmol/Lja- nus kinase 2/transcriptional activator 3 (JAK2/STAT3) pathway inhibitor AG490］and activator group ( 12.50 μmol/L ginkgolide B+0.50 μmol/L JAK2/STAT3 pathway activator Colivelin), 24 h after intervention, cell count kit 8 (CCK-8), 5-acetyl-2' deoxy- uridine (EdU) method, Hoechst 33258 staining, real-time fluorescence quantitative PCR (RT-qPCR) and protein immunoblot (WB) were used to detect cell viability, proliferation rate, apoptosis rate and expression levels of related factors. The results show that: compared with the control group, the cell viability ofthe medium/high dose test group and positive drug group was decreased (P <0.05), therefore, in this study, 12.50 μmol/L ginkgolide B group with significant difference and lower concentration was se- lected as ginkgolide B group for follow-up tests. Compared with the control group, the cell proliferation rate, Cyclin D1 mRNA and protein expression levels, p-JAK2 and p-STAT3 protein expression levels ofginkgolide B group and positive drug group were decreased, while the apoptosis rate and Caspase-3 mRNA and protein expression levels were increased (P <0.05). Compared with the ginkgolide B group, the changes of all indexes in inhibitor group were further enhanced (P <0.05). while those in activator group were significantly reversed (P <0.05). Ginkgolide B can inhibit the proliferation of OE19 cells and promote apoptosis of esophageal cancer cells by down-regulating JAK2/STAT3 signaling pathway. This study revealed a new anticancer mechanism of ginkgolide B and provided a theoretical basis for the study of esophageal cancer.