Abstract: Objective To study the effect of bufalin on apoptosis of human esophageal cancer ECA109 cells and its mechanism. Methods Inhibitory effect of bufalin on proliferation of human esophageal cancer ECA109 cells was assayed by MTT.Apoptosis of human esophageal cancer ECA109 cells and expression of survivin and caspase-3 were detected by flow cytometry and Western blot,respectively. Results The survival rate of human esophageal cancer ECA109 cells decreased while the apoptotic rate of human esophageal cancer ECA109 cells increased with the increasing concentration and time of bufalin used（P＜0.05）.The expression level of survivin and the activation caspase-3 protein in human esophageal cancer ECA109 cells increased with the increasing concentration of bufalin（P＜0.05）. Conclusion Bufalin induces apoptosis of human esophageal cancer ECA109 cells possibly by down-regulating the expression of survivin and the activation of caspase-3.