Brain-derived neurotrophic factor (BDNF) signals through its receptor tyrosine kinase (Trk)B to regulate the development trophoblast cells during peri- and postimplantation periods. Possible involvement of this signaling system in malignant human trophoblastic cell growth has not been investigated. Here, we found the expression of BDNF and neurtropin-4/5 together with TrkB in human trophoblastic choriocarcinoma cells. Treatment of cultured choriocarcinoma cells with a soluble TrkB ectodomain or a Trk receptor inhibitor K252a suppressed cell proliferation and increased apoptosis associated by the disruption of mitochondrial functions, whereas an inactive plasma membrane nonpermeable K252b was ineffective. Studies using these specific inhibitors also indicated the importance of the phosphatidylinositol 3-kinase and ERK pathways in mediating BDNF actions. Based on PCR array analyses to identify changes in expression profiles of cell cycle- and apoptosis-related genes in cultured choriocarcinoma cells, we found that suppression of endogenous TrkB signaling led to decreases in key proproliferation cell cycle genes and increases in two inhibitory cell cycle genes together with the up-regulation of several proapoptotic genes. In vivo studies in athymic nude mice bearing choriocarcinoma cell tumors further demonstrated that treatment with K252a, but not K252b, suppressed tumor growth accompanied by decreased cell proliferation, reduced levels of a tumor marker, human chorionic gonadotropin-beta, and increased levels of apoptosis and caspase-3/7 activities. Thus, autocrine signaling of the BDNF/TrkB system is important for human choriocarcinoma cell growth, and inhibition of BDNF/TrkB signaling in these cells could provide a novel therapy for patients with choriocarcinoma.