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Simple Summary Elevated PSA levels in blood tests are the gold standard for early prostate cancer detection, but its lack of specificity limits its clinical use as a mass screening test. The paradox is that it has long been known that advanced prostate cancers can lose PSA expression. We have observed that in the presence of tumors, the prostate produces and secretes less PSA than in healthy or benign conditions. Therefore, the PSA evaluation in urine provided more accurate information on the presence of prostate tumors than the blood test, representing a new method for the screening of prostate cancer. Abstract Serum prostatic specific antigen (PSA) has proven to have limited accuracy in early diagnosis and in making clinical decisions about different therapies for prostate cancer (PCa). This is partially due to the fact that an increase in PSA in the blood is due to the compromised architecture of the prostate, which is only observed in advanced cancer. On the contrary, PSA observed in the urine (uPSA) reflects the quantity produced by the prostate, and therefore can give more information about the presence of disease. We enrolled 574 men scheduled for prostate biopsy at the urology clinic, and levels of uPSA were evaluated. uPSA levels resulted lower among subjects with PCa when compared to patients with negative biopsies. An indirect correlation was observed between uPSA amount and the stage of disease. Loss of expression of PSA appears as a characteristic of prostate cancer development and its evaluation in urine represents an interesting approach for the early detection of the disease and the stratification of patients.