Protein synthesis plays a central role in cancer development and progression. eIF5A (eukaryotic initiation factor 5A), a translation factor activated by hypusination, is implicated in tumorigenesis, however, its mode of action is still unclear. Hypusinated eIF5A (eIF5Ahyp) promotes metastasis and tumor growth in prostate cancer (PCa) by supporting mitochondrial metabolism and translation. To characterize the pro-tumorigenic role of eiF5Ahyp in PCa, we employed pharmacological inhibition of deoxyhypusine synthase (DHPS) with GC7 in two PCa cell lines (LNCaP and DU145) coupled with quantitative proteomics.