Objective To investigate the prognosis of serum bicorticotin-like kinase 1 （DCLK1） and soluble T cell immunoglobulin mucin molecule 3 （sTim-3） in patients undergoing radical prostatectomy. Methods This study selected 80 patients who underwent radical prostatectomy in our hospital from June 2016 to June 2019 as the research group， and another 80 healthy individuals who underwent physical examination during the same period as the control group； enzyme linked immunosorbent assay （ELISA） was applied to detect serum levels of DCLK1 and sTim-3； The relationship between serum DCLK1 and sTim-3 and pathologic features and prognosis of patients undergoing radical prostatectomy was analyzed. Results The serum levels of DCLK1 and sTim-3 in the study group were obviously higher than those in the control group （P < 0.05）. Serum DCLK1 and sTim-3 levels were positively correlated （P < 0.05）. Both were positively correlated with PSA level and Gleason score （P < 0.05）.Serum DCLK1 and sTim-3 expression levels were associated with PSA levels， Gleason scores， and TNM stages （P < 0.05）. The serum levels of DCLK1 and sTim-3 in the prognostic death group were significantly higher than those in the prognostic survival group （P < 0.05）. Patients with high expression of DCLK1 and sTim-3 had lower survival rates than those with low expression. According to Cox regression analysis， high expression of DCLK1 and sTim-3 was a risk factor affecting the prognosis of patients undergoing radical prostatectomy （P < 0.05）. According to the ROC curve， the AUC that jointly predicted the prognosis of patients undergoing radical prostatectomy was better than that of DCLK1 and sTim-3 respectively. Conclusion DCLK1 and sTim-3 are obviously elevated in the serum of prostate cancer patients， and their combination can better evaluate the prognosis of patients undergoing radical prostatectomy.