As a novel mode of tumor neovascularization,vasculogenic mimicry (VM) has been reported to increase tumor-related mortality in many different solid tumors. In the present study,two established human gallbladder carcinoma (GBC) cell lines ( highly aggressive GBC-SD and poorly aggressive SGC-996) cultured on a three-dimensional matrix were assessed for the ability of VM channel formation under normoxic or hypoxic conditions. In addition,the relationship between HIF-1α gene expression and VM channel formation of GBC cells in vitro was measured by using the small interfering RNA (siRNA) technique,western blotting and real-time reverse transcription (RT) -PCR analysis. Furthermore,H&E and CD31/periodic acid-Schiff (PAS) staining were used to observe VM in GBC tissue samples. Additionally,all seventy-one specimens with VM and non-VM were stained for hypoxia inducible factor-1α (HIF-1α) and its correlation with clinicopathological features and prognosis was analyzed simultaneously . We found that hypoxia could induce more VM channel formation and elevated HIF-1α expression in highly aggressive GBC-SD cells. HIF-1α siRNA efficiently knocked down HIF-1α expression and GBC VM networks under either normoxic or hypoxic conditions. VM was present in human primary GBC and overexpression of HIF-1α was significantly correlated with depth of invasion and perineural involvement in the non-VM group. Moreover,VM and HIF-1α were independent factors for the overall survival of GBC patients and correlated with decreased survival. In conclusion,VM was present in human GBC. As a critical mediator in VM formation,high expression of HIF-1α was associated with VM and tumor progression in GBC patients. As a novel mode of tumor neovascularization, vasculogenic mimicry (VM) has been reported to increase tumor-related mortality in many different solid tumors. In the present study, two established human gallbladder carcinoma (GBC) cell lines poorly aggressive SGC-996) cultured on a three-dimensional matrix were assessed for the ability of VM channel formation under normoxic or hypoxic conditions. In addition, the relationship between HIF-1α gene expression and VM channel formation in GBC cells in vitro was measured by using the small interfering RNA (siRNA) technique, western blotting and real-time reverse transcription (RT) -PCR analysis. Further, H & E and CD31 / periodic acid-Schiff Additionally, all seventy-one specimens with VM and non-VM were stained for hypoxia inducible factor-1α (HIF-1α) and its correlation with clinicopathological features and prognosis was yet analyzed simultaneously. We fou nd that hypoxia could induce more VM channel formation and elevated HIF-1α expression in highly aggressive GBC-SD cells. HIF-1α siRNA efficiently knocked down HIF-1α expression and GBC VM networks either either normoxic or hypoxic conditions. VM was present in human Primary GBC and overexpression of HIF-1α were significantly correlated with depth of invasion and perineural involvement in the non-VM group. Moreover, VM and HIF-1α were independent factors for the overall survival of GBC patients and correlated with decreased survival. , VM was present in human GBC. As a critical mediator in VM formation, high expression of HIF-1α was associated with VM and tumor progression in GBC patients.