Prostate cancer is initially treated via androgen deprivation therapy (ADT), a highly successful treatment in the initial pursuit of tumor regression, but commonly restricted by the eventual emergence of a more lethal 'castrate resistant' form of the disease. Intracrine pathways that utilize dehydroepiandrosterone (DHEA) or other circulatory precursor steroids, are thought to generate relevant levels of growth-stimulating androgens such as testosterone (T) and dihydrotestosterone (DHT). In this study, we explored the capacity of the active vitamin D hormone to interact and elicit changes upon this prostatic intracrine pathway at a metabolic level. We used androgen dependent LNCaP cells cultured under steroid-depleted conditions and assessed the impact of vitamin D-based compounds upon intracrine pathways that convert exogenously added DHEA to relevant metabolites, through Mass Spectrometry (MS). Changes in relevant metabolism-related gene targets were also assessed. Our findings confirm that exposure to vitamin D based compounds, within LNCaP cells, elicits measurable and significant reduction in the intracrine conversion of DHEA to T, DHT and other intermediate metabolites within the androgenic pathway. The aassessment and validation of the biological model and analytical platforms were performed by pharmacological manipulations of the SRDS alpha and HSD-17 beta enzymes. The data provides further confirmation for how a vitamin D-based regime may be used to counter intracrine mechanisms contributing to the emergence of castrate-resistant tumors. (C) 2019 Elsevier Inc. All rights reserved.