Abstract: Objective To investigate hypoxic microenvironment on esophageal cancer cells Ec9706 vasculogenic mimicry and its possible molecular mechanism. Methods Human esophageal cancer cells were induced by CoCl2 chemical method to form hypoxia-microenvironment. While in hypoxic microenvironment, human esophageal squamous cell carcinoma cell lines Ec9706 was interfered by Bcl-2-siRNA, and human body environment was simulated by three-dimensional culture system and placed in the inverted fluorescence microscope to observe the number of pipeline structure formation and morphology. Flow cytometry was applied to detect cell proliferation. RT-PCR and Western blot were used to assay mRNA and protein expression of Bcl-2, VE-cad and MMP2. Results Hypoxia group formed more network-like tubular structure (P<0.000) than normoxic control group, decreased apoptosis (P<0.05), the mRNA and protein expression of Bcl-2, VE-cad and MMP2 compared with normoxic significantly increased (P<0.05) lower. After training in hypoxia carried out under Bcl-2-siRNA interference, the tubular network-like structure of experimental group significantly reduced than hypoxic hypoxia control group (P<0.000), apoptosis increased (P<0.05), whereas the mRNA and protein expression of Bcl-2, VE-cad and MMP2 was also significantly lower (P<0.05). The network-like tubular structures between hypoxia and anoxia empty vector control was no significant difference (P>0.05), apoptosis rate was no statistically significant (P>0.05), the mRNA and protein of Bcl-2, VE-cad and MMP2 was not statistically significant between groups(P>0.05). Conclusion Hypoxia could induce the formation of esophageal cancer cells Ec9706 vasculogenic mimicry, and the important molecular mechanism may be related to Bcl-2-dependent VE-cad overexpression.