AimTo investigate the association between positive lesions detected by 99mTc-PSMA SPECT/CT and blood levels of prostate-specific antigen (PSA) and alkaline phosphatase (ALP) in patients with prostate cancer (PCa) and bone metastasis undergoing endocrine therapy. MethodsA retrospective analysis was performed on 43 patients diagnosed with PCa bone metastasis who underwent endocrine therapy. PSA, ALP, whole body bone imaging and 99mTc-PSMA SPECT/CT imaging were collected from all patients (Among them, 17 cases were re-examined 99mTc-PSMA SPECT/CT imaging). According to the results of the first 99mTc-PSMA SPECT/CT imaging for detecting bone metastasis, all cases were divided into two groups: positive group and negative group. The relationship between 99mTc-PSMA imaging and PSA and ALP was analyzed by ROC curve. Fisher exact probability method was used to examine the changes in imaging radioactivity uptake, PSA, and ALP levels in 17 patients after treatment, and P < 0.05 was statistically significant. ResultsAll 43 patients had different degrees of radioactive concentrations on whole-body bone imaging. The first 99mTc-PSMA SPECT/CT imaging showed positive bone metastases in 31 cases and negative bone metastases in 12 cases. ROC curve analysis of PSA and ALP, AUC were 0.778 and 0.770, respectively. When PSA > 1.13 ng/mL, 99mTc-PSMA SPECT/CT imaging diagnostic sensitivity was 93.55%, and specificity was 66.67%. When ALP was >86U/L, the diagnostic sensitivity of 99mTc-PSMA SPECT/CT imaging was 64.52%, and the specificity was 83.33%. In 17 cases, the PSA level decreased in 7 and increased in 10. There were 10 cases of increased ALP and 7 cases of decreased ALP levels. In the second 99mTc-PSMA imaging lesion, there were 9 cases with decreased or no uptake, and 8 cases with increased uptake or number of lesions. The changes in 99mTc-PSMA uptake by Fisher’s exact probability method were statistically significant (P < 0.05, P = 0.006, and P = 0.006, respectively), and ALP level was not statistically significant (P = 0.563). Conclusion99mTc-PSMA SPECT/CT imaging can detect PCa bone metastases, which are related to PSA levels. When PSA > 1.13 ng/mL, the sensitivity of diagnosis and detection of positive bone metastases is higher, and when ALP is >86U/L, 99mTc-PSMA imaging has higher specificity.