Research over the past decade has identified a number of biomarkers that are associated with high Gleason grade disease. Previous studies from our laboratory found a correlation between expression of FLICE-inhibitory protein and tumor grade in human prostate cancer. We further demonstrated that Sp1 transactivates the FLIP promoter while Sp3 inhibits Sp1-mediated trans-activation, thus implicating a role for these factors during prostate carcinogenesis. However, it was not known whether any of these markers could achieve the sensitivity and specificity necessary to distinguish aggressive from indolent disease. Here, we evaluated whether the “biomarker signature” of FLIP, Sp1, and Sp3 can predict the development of prostate cancer recurrence by immunohistochemical evaluation of tissue samples obtained from patients who underwent prostatectomy as primary treatment for prostate cancer and were observed for at least 5 years with PSA measurements. We show that the combination of FLIP, Sp1, Sp3, and Gleason score is an excellent predictor of biochemical recurrence. The area under the receiver operator characteristic curve for FLIP, Sp1, and Sp3 when predicting PSA failure was 0.71, 0.66, and 0.68 respectively; however, when these three markers were combined with Gleason score the AUC increased to 0.93. This level of prediction for PSA failure suggests that this biomarker panel could be an important predictor of biochemical recurrence. Effective clinical management of PCA has been hampered by TWS119 significant intratumoral heterogeneity combined with an incomplete understanding of the molecular events associated with the development of the disease and subsequent recurrence following traditional treatments. Therefore, there is an unmet need for new methods and/or agents for PCA management. Given the individual genetic variation and the heterogeneity of the disease, personalized treatment approaches are critical for successful management of PCA. To develop such individualized treatment approaches, it is essential to identify a panel of biomarkers or a “biomarker signature” that could be used to stratify patients according to response to specific treatments. To the best of our knowledge, there are currently no sensitive markers to monitor disease recurrence. In this study we assessed the expression of the anti-apoptotic protein FLIP and the transcription factors Sp1 and Sp3 by immunohistochemical evaluation of tissue samples obtained from 64 patients who underwent radical prostatectomy as primary treatment for prostate cancer. We believe that this is the first report of FLIP, Sp1, and Sp3 expression and the correlation among these proteins in biochemically recurrent PCA samples. Although increased expression of Sp1, Sp3, or FLIP showed significant differences between PSA failure and non-failure cases, individually they are not strong predictors of poor clinical outcome based on AUC when PSA failure is used as a surrogate outcome: the area under the ROC curve for FLIP, Sp1, Sp3, and Gleason as a predictor of PSA failure and non-failure cases was 0.71, 0.66, 0.68, and 0.76 respectively. On the other hand, the biomarker signature of Sp1/Sp3/FLIP combined with Gleason achieved an AUC of 0.93. These data indicate excellent discrimination between PSA failure and non-failure cases and suggest that this biomarker signature is an important predictor of the probability of biochemical recurrence.