Objective To explore the mechanism of nodakenin (NDK) affecting the invasion and migration of cervical cancer cells by regulating epithelial-mesenchymal transformation (EMT), and analyze its effect on microRNA (miR)-324-3P/WNT3a axis. Methods Human cervical cancer HeLa cells in logarithmic growth phase were divided into four groups: blank group(cultivated in a 37 ℃, 5% CO₂-saturated humidity incubator), NDK group (20 mmol/L NDK was added to culture), miR-324-3P inhibitor group (hsa-miR-324-3p inhibitor transfection), and miR-324-3P inhibitor+NDK group (after transfection of miR-324-3P inhibitor, 20 mmol/L NDK was added to culture) all were treated for 48 h. Transwell test was used to detect the invasion ability of cells in each group. Cell scratch test was performed to detect cell migration ability of each group. qRT-PCR to detect the mRNA level of miR-324-3P and WNT3a in each group of cells. Western blotting was carried out to detect the expression of EMT-related marker proteins E-cadherin, vimentin, Neural cadherin (N-cadherin), WNT3a, and β-catenin protein in each group of cells. Dual luciferase reporter gene was used to detect the targeting relationship between miR-324-3P and WNT3a gene. Results Compared with the blank group, the number of transmembrane HeLa cells increased in miR-324-3P inhibitor group, the scratch healing rate increased, the mRNA expression level of miR-324-3P decreased, and of WNT3a increased; the relative expression of E-cadherin protein decreased, while the relative expressions of vimentin, N-cadherin, WNT3a and β-catenin increased (P<0.05). While for NDK group and miR-324-3P inhibitor+NDK group, the number of transmembrane HeLa cells decreased, the scar healing rate was also decreased, the mRNA expression level of miR-324-3P increased, the expression level of WNT3a mRNA decreased, the relative expression of E-cadherin protein increased, and the relative expression of vimentin, N-cadherin, WNT3a and β-catenin protein decreased (P<0.05); All the indexes mentioned above were lower in NDK group than those in miR-324-3P inhibitor+NDK group.The dual luciferase reporter gene results showed that miR-324-3P mimic could reduce WNT3a WT luciferase activity (P<0.05), but did not affect WNT3a mut luciferase activity (P>0.05). Conclusion NDK can inhibit the invasion and migration of cervical cancer cells by regulating EMT, and the mechanism of such action may be related to the regulation of miR-324-3P/WNT3a axis.