ABSTRACT: Purpose: The purpose of this study was to investigate the feasibility of Single-Photon Emission Computed Tomography/Computed Tomography (SPECT/CT) image-based radiomics in differentiating bone metastases from benign bone lesions in patients with tumors. Methods: A total of 192 lesions from 132 patients (134 in the training group, 58 in the validation group) diagnosed with vertebral bone metastases or benign bone lesions were enrolled. All images were evaluated and diagnosed independently by two physicians with more than 20 years of diagnostic experience for qualitative classification, the images were imported into MaZda software in Bitmap (BMP) format for feature extraction. All radiomics features were selected by least absolute shrinkage and selection operator (LASSO) regression and 10-fold cross-validation algorithms after the process of normalization and correlation analysis. Based on these selected features, two models were established: The CT model and SPECT model (radiomics features were derived from CT and SPECT images, respectively). In addition, a combination model (ComModel) combined CT and SPECT features was developed in order to better evaluate the predictive performance of radiomics models. Subsequently, the diagnostic performance between each model was separately evaluated by a confusion matrix. Results: There were 12, 13, and 18 features contained within the CT, SPECT, and ComModel, respectively. The constructed radiomics models based on SPECT/CT images to discriminate between bone metastases and benign bone lesions not only had high diagnostic efficacy in the training group (AUC of 0.894, 0.914, 0.951 for CT model, SPECT model, and ComModel, respectively), but also performed well in the validation group (AUC; 0.844, 0.871, 0.926). The AUC value of the human experts was 0.849 and 0.839 in the training and validation groups, respectively. Furthermore, both SPECT model and ComModel show higher classification performance than human experts in the training group (P = 0.021 and P = 0.001, respectively) and the validation group (P = 0.037 and P = 0.007, respectively). All models showed better diagnostic accuracy than human experts in the training group and the validation group. Conclusion: Radiomics derived from SPECT/CT images could effectively discriminate between bone metastases and benign bone lesions. This technique may be a new non-invasive way to help prevent unnecessary delays in diagnosis and a potential contribution in disease staging and treatment planning.