Objectives: To study the effect of Tspan5 on the proliferation and migration of trophoblast cells. Materials and Methods: The authors silenced the Tspan5 expression in human choriocarcinoma cell line JEG-3 using RNAi, and then conducted the cell proliferation assay and scratch assay to detect the proliferation and migration of JEG-3 cells. Results: The present authors found that the JEG-3 cell proliferation and migration activity of interference group (Tspan5/1005) had decreased significantly. Tspan5/1005 moderately decreased FAK and AKT phosphorylation compared to the control group. In this study, the authors found that the proliferation and migration of choriocarcinoma cells were reduced after using RNAi technology to downregulate the expression level of Tspan5 expression. The level of Tspan5 expression was positively correlated with the proliferation and migration of choriocarcinoma cells. Conclusions: Tspan5 could play a biological role through the FAK/AKT signaling pathway.