Objective: To develop a machine learning (ML)-assisted model capable of accurately predicting the probability of biopsy Gleason grade group upgrading before making treatment decisions by integrating multiple clinical characteristics.Materials and Methods: We retrospectively collected data from PCa (prostate cancer) patients who underwent systematic biopsy and radical prostatectomy from January 2015 to December 2019 at Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology. The study cohort was divided into training and testing datasets in a 70:30 ratio for further analysis. Four ML-assisted models were developed from 16 clinical features using logistic regression (LR), logistic regression optimized by Lasso regularization (Lasso-LR), random forest (RF) and support vector machine (SVM). The area under the curve (AUC) was applied to determine the model with the highest discrimination. Calibration plots were used to investigate the extent of over- or underestimation of predicted probabilities relative to the observed probabilities in models. Results: In total, 530 PCa patients were included, with 371 patients in the training dataset and 159 patients in the testing dataset. The Lasso-LR model showed good discrimination with an AUC, accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 0.776, 0.712, 0.679, 0.745, 0.730 and 0.695, respectively, followed by SVM (AUC 0.740, 95%CI: 0.690–0.790), LR (AUC 0.725, 95%CI: 0.674–0.776) and RF (AUC 0.666, 95%CI: 0.618–0.714). Validation of the model showed that the Lasso-LR model had the best discriminative power (AUC 0.735, 95%CI: 0.656–0.813), followed by SVM (AUC 0.723, 95%CI: 0.644–0.802), LR (AUC 0.697, 95%CI: 0.615–0.778) and RF (AUC 0.607, 95%CI: 0.531–0.684) in the testing dataset. Both the Lasso-LR and SVM models were well-calibrated. Conclusions: The Lasso-LR model had good discrimination in the prediction of patients at high risk of harboring incorrect Gleason grade group assignment, and the use of this model may be greatly beneficial to urologists in treatment planning, patient selection, and the decision-making process for PCa patients.