Over‐expression of hexose transporters (Gluts), specifically Glut‐1, is a common event in human malignancies. In prostate cancer (CaP), however, expression of Gluts has been characterized poorly. In this study, expression and distribution of Glut‐1 and Glut‐5 proteins were characterized using immunohistochemistry in 76 specimens of benign prostate, 10 specimens of high‐grade intraepithelial neoplasia (HGPIN), and 28 specimens of CaP. In addition, mRNA expression of Glut‐2, Glut‐7, Glut‐9, and Glut‐11 was analyzed in a set of five specimens of benign prostate and CaP. In benign prostate, Glut‐1 localized to the basal cells and to the basolateral membrane of secretory/luminal epithelial cells. Glut‐5, however, localized to the apical membrane of secretory/luminal epithelial cells. In HGPIN, Glut‐1 was immunohistochemically undetectable. Glut‐5, however, localized to the apical membrane of the neoplastic epithelial cells. In CaP, Glut‐1 and Glut‐5, were immunohistochemically undetectable. However, over‐expression of GLUT1 was observed in some specimens of highly proliferative intraductal CaP. Glut‐7, Glut‐9, and Glut‐11 mRNAs were detected in benign prostate and CaP, however, only Glut‐11 mRNA was consistently up‐regulated in CaP compared to benign prostate. Low levels of expression of Glut‐1 protein in the majority of CaP could explain, at least in part, the limited clinical applicability of positron emission tomography using 2‐[18F]‐fluoro‐2‐deoxy‐D‐glucose for imaging CaP. Moreover, expression of Glut‐5 in HGPIN suggested that fructose could be utilized as potential metabolic substrate in HGPIN. Understanding the molecular mechanisms involved in regulation/dysregulation of Gluts in CaP could provide insight in the understanding of hexose metabolism in CaP. J. Cell. Biochem. 113: 553–562, 2012. © 2011 Wiley Periodicals, Inc.