Background-Estrogens may induce the proliferation of neoplastic cells by activating neoangiogenesis.Aim-To evaluate the effect of estrogens on the expression of vascular endothelial growth factor (VEGF) and related receptors (VEGF-R) in human cholangiocarcinoma and the role played by VEGF in mediating the proliferative effects of estrogens.Methods-Seven biopsies of intra-hepatic cholangiocarcinoma and the HuH-28 cell lines were investigated. Cell proliferation was measured by both PCNA Western blot and MTS proliferation assay.Results-By immunohistochemistry, biopsies of human cholangiocarcinoma stained positively for VEGF-A and VEGF-C and related receptors. HuH-28 cells expressed VEGF-A, -C, and VEGFR-1, -2, -3 and, their protein level was enhanced by 17β-estradiol in association with the stimulation of cell proliferation. 17β-Estradiol-stimulated proliferationof HuH-28 cells was blocked by 70% by VEGF-trap, a receptor-based VEGF inhibitor. 17β-Estradiol induced the secretion of VEGF in the