The purpose of this study was to retrospectively characterize benign and malignant prostate peripheral zone tissue by using endorectal MRI and 3D ¹H MRS. Fifty-two men with untreated biopsy-proven prostate cancer underwent combined endorectal MRI and MRSI. Whole-mount step-section histopathologic analysis constituted the reference standard. Biopsy correctly detected 74 locations; MRI correctly detected 72 locations; MRS correctly detected 72 locations; MRI + MRS correctly detected 78 locations. Cohen's test showed that biopsy had a lower degree of agreement with histology than MRI + MRS combined. The ratio of [(Cho + Cr)]/Cit correlates with the pathologic Gleason score. The addition of 3D¹H MRSI to MRI can improve diagnosis of prostate cancer contributing indirectly to improve local staging. In addition, the correlation between metabolic 3D¹H MRSI data with pathological Gleason grade may offer a non-invasive means to better predict prostate cancer aggressiveness.