Understanding the links between obesity and prostate cancer will have major implications for the health policy for men with prostate cancer and the development of new therapeutic or preventative strategies. Cancer metabolism is a hallmark of cancer pathogenesis and is required to support the malignant properties of cancer cells, particularly in endocrine-related cancers such as breast and prostate cancer. This dysregulation is exacerbated in obesity, where patients develop excess adipose tissue with dysfunctional lipid metabolism and endocrine function that impacts on cancer cell survival. Previous studies have focused extensively on the roles of glucose, glutamate and fatty acids derived from de novo lipogenesis in modulating the bioenergetic processes and macromolecule synthesis required to sustain growth and proliferation. However, fatty acids are also derived from adipose tissue lipolysis or the breakdown of triglycerides contained in circulating chylomicrons and lipoproteins. We investigated the role of lipid metabolism in prostate cancer using tissue from patients with prostate cancer and patient-derived xenograft mouse models. We showed that fatty acid uptake was increased in human prostate cancer and that these fatty acids were directed toward biomass production. These changes were mediated, at least partly, by the fatty acid transporter CD36, which was associated with aggressive disease. Deleting Cd36 in the prostate of cancer-susceptible Pten−/− mice reduced fatty acid uptake and the abundance of oncogenic signaling lipids and slowed cancer progression. Moreover, CD36 antibody therapy reduced cancer severity in patient-derived xenografts. We further demonstrated cross-talk between fatty acid uptake and de novo lipogenesis and found that dual targeting of these pathways more potently inhibited proliferation of human cancer-derived organoids compared to the single treatments. These findings identify a critical role for CD36-mediated fatty acid uptake in prostate cancer and suggest that targeting fatty acid uptake might be an effective strategy for treating prostate cancer.