Objective To investigate the clinical application value of three-dimensional ultrasonography and color Doppler ultrasonography in the evaluation of neoadjuvant chemotherapy for breast cancer. Methods In this study 80 breast cancer patients confirmed by pathology who received neoadjuvant chemotherapy in Xiangya Hospital of Central South University were collected. Before and after neoadjuvant chemotherapy, all the patients underwent clinical assessment, two-dimensional (2D) ultrasonographic and three-dimensional (3D) ultrasonographic examinations. The results were compared with the pathological evaluation results according to the changes of breast masses. Results There was a significant difference between clinical assessment and pathological assessment (P < 0.05). However, the 2D ultrasonographic and 3D ultrasonographic assessments were not significantly different from the pathological assessment (P > 0.05). The maximum diameters of the residual tumors after neoadjuvant chemotherapy by 2D and 3D ultrasound measurement were positively correlated with those by pathological evaluation (P < 0.05). In the effective group, the differences in the dropping grade of blood flow grading and reduction of blood flow resistance index before and after chemotherapy had statistical significance (P < 0.05). Conclusions 3D ultrasonographic assessment has high consistency with pathological evaluation for curative effect of neoadjuvant chemotherapy. The change of ultrasound blood flow signal in breast cancer masses can be helpful to determine the curative effect of neoadjuvant chemotherapy.