Objective To explore the effect of Netrin-1 on the induction of cervical cancer epithelial-mesenchymal transition (EMT) by activating the PI3K/AKT pathway under hypoxic conditions. Methods The expression of Netrin-1 in normal HaCaT epithelial cells and SiHa, Caski and Hela cervical cancer cell lines were detected. The cobalt dioxide treatment simulated the hypoxic microenvironment. The invasive and migratory ability of SiHa, Caski and Hela cells were detected through Transwell assay and cell wound healing assay. The expression of Netrin-1and EMT markers (E-cadherin and Vimentin) was examined using Western Blotting. SiHa cells were transfected with small interfering RNA (Si-Netrin-1) to detect the expression of E-cadherin, Vimentin, AKT, p-AKT proteins and the changes in cell invasive and migratory capabilities. Results Netrin-1 was highly expressed in cervical cancer cell lines, and the expression of Netrin-1 was significantly enhanced under hypoxic conditions (P＜0.01). E-cadherin was significantly down-regulated and Vimentin was up-regulated in SiHa and Caski cells under hypoxic conditions, cell invasive and migratory capabilities were also significantly enhanced, in addition, PI3K/AKT signaling pathway was activated. Netrin-1 knockdown inhibited EMT of SiHa cells induced by hypoxia. Conclusion Netrin-1 induces epithelial-mesenchymal transition of cervical cancer through activating PI3K/AKT signaling pathway under hypoxic conditions.