Abstract: OBJECTIVE To investigate the effects and the related molecular mechanisms of cordycepin on cell proliferation, motility and apoptosis of human gastric carcinoma cell lines HGC-27. METHODS Human gastric carcinoma cell lines HGC-27, AGS and MGC-803 were treated with different doses of cordycepin. Cell counting kit-8 assay was used to determine relative cell vitality. Cell proliferation ability was assessed by colony formation assay. Cell apoptosis was analyzed by flow cytometry and cell motility was detected by healing wound assay. Western blot was used to test the expression levels of Bcl-2, Bax, ERK, p-ERK and GSK-3β. RESULTS Compared with the control group, cordycepin treatment groups could significantly inhibit cell proliferation, the rate of colony formation and cell motility, induce cell apoptosis and down-regulate the expression levels of Bcl-2, GSK-3β and p-ERK in a dose and time-dependent manner, but it could not affect the expression level of Bax. CONCLUSION Cordycepin exert significant anticancer effects by modulating ERK/GSK-3β signaling pathway, which may be potential drug for cancer prevention and therapy of gastric carcinoma in the clinic.