Objective To investigate the application of CT image and cone beam computed tomography (CBCT) image registration based on 3D Slicer software in image-guided radiotherapy for uterine cervical neoplasms. Methods Based on 3D Slicer software and Slicer RT toolkit, 10 positioning CT images and 50 CBCT images of 10 patients with uterine cervical neoplasms in Henan Provincial People's Hospital between January 2018 and October 2018 had rigid registration and b-spline deformation registration respectively. The dice similarity coefficient (DSC) and Hausdorff distance (HD) of the bladder, rectum, femoral head, spinal cord, and body of CT-CBCT images were compared by using paired t-test before and after the registration. Results Pre-registration, rigid registration and after b-spline deformation registration of CT images and CBCT images, the DSC in the bladder (0.459±0.177, 0.528±0.184, 0.542±0.187, respectively), the rectum (0.564±0.141, 0.632±0.091, 0.684±0.097, respectively), the femoral head (0.695±0.088, 0.833±0.030, 0.865±0.027, respectively), the spinal cord (0.587±0.119, 0.746±0.085, 0.834±0.032, respectively) and the body surface (0.922±0.013, 0.948±0.011, 0.959±0.009, respectively) showed an increased trend; HD in the bladder (12.8±7.2, 12.2±7.1, 11.7±7.3, respectively), the rectum (5.0±1.8, 4.4±1.2, 3.4±1.2, respectively), the femoral head (3.6±1.2, 1.8±0.5, 1.5±0.5, respectively), the spinal cord (4.0±1.0, 2.7±1.3, 1.8±0.5, respectively) and the body surface (6.3±2.1, 5.2±2.0, 4.3±2.0, respectively) showed a decreased trend. The differences of pairwise comparison in the same parts were statistically significant (all P < 0.05). Conclusions Both rigid registration and b-spline deformation registration of CT-CBCT images based on 3D Slicer software can improve the radiotherapy accuracy of uterine cervical neoplasms, and b-spline deformation registration has more significant advantages. Key words: Uterine cervical neoplasms; Radiotherapy; 3D Slicer software; Cone-beam computed tomography; Image registration; Image-guided radiation therapy