Objective: To generate virtual non-contrast (VNC) computed tomography (CT) from intravenous enhanced CT through convolutional neural networks (CNN) and compare calculated dose among enhanced CT, VNC, and real non-contrast scanning. Method: 50 patients who accepted non-contrast and enhanced CT scanning before and after intravenous contrast agent injections were selected, and two sets of CT images were registered. A total of 40 and 10 groups were used as training and test datasets, respectively. The U-Net architecture was applied to learn the relationship between the enhanced and non-contrast CT. VNC images were generated in the test through the trained U-Net. The CT values of non-contrast, enhanced and VNC CT images were compared. The radiotherapy treatment plans for esophageal cancer were designed, and dose calculation was performed. Dose distributions in the three image sets were compared. Results: The mean absolute error of CT values between enhanced and non-contrast CT reached 32.3 ± 2.6 HU, and that between VNC and non-contrast CT totaled 6.7 ± 1.3 HU. The average CT values in enhanced CT of great vessels, heart, lungs, liver, and spinal cord were all significantly higher than those of non-contrast CT (p < 0.05), with the differences reaching 97, 83, 42, 40, and 10 HU, respectively. The average CT values of the organs in VNC CT showed no significant differences from those in non-contrast CT. The relative dose differences of the enhanced and non-contrast CT were −1.2, −1.3, −2.1, and −1.5% in the comparison of mean doses of planned target volume, heart, great vessels, and lungs, respectively. The mean dose calculated by VNC CT showed no significant difference from that by non-contrast CT. The average γ passing rate (2%, 2 mm) of VNC CT image was significantly higher than that of enhanced CT image (0.996 vs. 0.973, p < 0.05). Liugang et al. Non-contrast CT Generated From Enhanced Conclusion: Designing a treatment plan based on enhanced CT will enlarge the dose calculation uncertainty in radiotherapy. This paper proposed the generation of VNC CT images from enhanced CT images based on U-Net architecture. The dose calculated through VNC CT images was identical with that obtained through real non-contrast CT.