Objective: To investigate the role and the correlation of HIF-1α and EphA2 in the process of vasculogenic mimicry in esophageal squamous cell carcinoma. Methods: Two esophageal cancer cell lines Eca109 and TE13 were transfected by plasmid harboring interfering RNA targeting HIF-1α and EphA2. The relationship of the expression of HIF-1α and EphA2 was measured by RT-PCR and Western blot. The formation of tubular networks of Eca109 and TE13 cells was analyzed by three-dimension culture in vitro.Results: ①The expression of EphA2 in Eca109 and TE13 was inhibited with silencing HIF-1α. On the contrary, the expression of HIF-1α in Eca109 and TE13 was not obviously inhibited with silencing EphA2. ②The mRNA expression of HIF-1α and EphA2 were markedly inhibited in Eca109 and TE13 lines with silencing HIF-1α and EphA2 gene. ③The numbers of tubular networks were both remarkably decreased in Eca109 and TE13 with silencing HIF-1α and EphA2 gene, but the number of the former was more than the latter. Conclusion: HIF-1α and EphA2 may both participate the formation of vasculogenic mimicry of esophageal squamous cell carcinoma. EphA2 may be one of the important factors regulated by HIF-1αin the process.