Objective To investigate the effect and mechanism of erythropoietin producing hepatoma receptor (Eph) receptor A2 gene silencing on migration and invasion of human esophageal cancer EC9706 cells.Methods Human esophageal cancer EC9706 cells in logarithmic growth phase were randomly divided into control group, empty group, and silent group, transfected with Lipofectamine ^TM 2000, containing nonsense random sequence (siRNA-NC-PGCsi3.0) and EphA2 interference sequence (EphA2-siRNA-PGCsi3.0). After stable transfection, Brd U method was used to detect cell proliferation ability, scratch test to detect cell migration ability, Transwell cell test to detect invasion ability, qRT-PCR and Western blotting to detect the mRNA and protein expression levels of EphA2, Wnt1, β-catenin, E-cadherin, Vimentin in cells.Results The relative expression of EphA2 mRNA in the control group, no-load group, and silent group were (1.78 ± 0.17), (1.82 ± 0.18), (0.37 ± 0.11), respectively. The difference between the groups was statistically significant (P < 0.05). Compared with the control group and the no-load group, the relative expression of EphA2 mRNA in the silent group decreased (P < 0.05). The positive rates of Brd U in the control group, the no-load group, and the silent group were (26.48 ± 2.79) %, (23.52 ± 2.57) %, (13.29 ± 2.06) %, respectively. The difference between the groups was statistically significant (P < 0.05). Compared with the control group and the no-load group, the positive rate of Brd U in the silent group was lower (P < 0.05). The cell scratch healing rates of the control group, no-load group, and silent group were (83.16 ± 8.31) %, (82.35 ±7.24) %, (34.24 ± 5.27) %, respectively. The difference between the groups was statistically significant (P < 0.05). Compared with the control group and the no-load group, the cell scratch healing rate of the silent group increased (P < 0.05). The number of transmembrane cells in the control group, the no-load group, and the silent group were (326.74 ± 33.15), (331.27 ± 34.59), (126.23 ± 26.18), respectively. The difference between the groups was statistically significant (P < 0.05). Compared with the control group and the no-load group, the number of transmembrane cells in the silent group decreased (P < 0.05). The differences of relative expression levels of Wnt1, β-catenin, E-cadherin, and Vimentin mRNA were statistically significant (P < 0.05). Compared with the control group and the no-load group, the relative expression of Wnt1, β-catenin, Vimentin mRNA in the silent group decreased, and the relative expression of E-cadherin mRNA increased (P < 0.05). The difference of relative expression levels of EphA2, Wnt1, β-catenin, E-cadherin, and Vimentin protein in cells were statistically significant (P < 0.05). Compared with the control group and the no-load group, the relative expression of EphA2, Wnt1, β-catenin, and Vimentin protein in the silent group decreased, and the relative expression of E-cadherin protein increased (P < 0.05). The comparison of the above indicators between the control group and the no-load group showed no significant difference (P > 0.05).Conclusion EphA2 gene silencing has a certain inhibitory effect on the migration and invasion of human esophageal cancer EC9706 cells, and its mechanism may be related to preventing the Wnt/β-catenin signaling pathway.