Cancer etiology is influenced by alterations in protein synthesis which are not fully understood. In this study, we took a novel approach to investigate the role of the eukaryotic translation initiation factor eIF5A in human cervical cancers where it is widely overexpressed. eIF5A contains the distinctive amino acid hypusine, which is formed by a post-translational modification event requiring deoxyhypusine hydroxylase (DOHH), an enzyme that can be inhibited by the drugs ciclopirox and deferiprone. We found that proliferation of cervical cancer cells could be blocked by DOHH inhibition with either of these pharmacological agents, as well as by RNAi-mediated silencing of eIF5A, DOHH or other enzymes in the hypusine pathway. Proteomic and RNA analyses in HeLa cervical cancer cells identified two groups of proteins in addition to eIF5A that were coordinately affected by ciclopirox and deferiprone. Group 1 proteins (HSP27, NM23 and DJ1) were downregulated at the translational level, whereas Group 2 proteins (TrpRS and PRDX2) were upregulated at the mRNA level. Further investigations of Hsp27 control confirmed that eIF5A and DOHH were required for its expression in cervical cancer cells and for regulation of its key targets I?B and NF?B there. Our results argue that mature eIF5A controls a translational network of cancer-driving genes, termed the eIF5A regulon, at the levels of mRNA abundance and translation. In coordinating cell proliferation, the eIF5A regulon can be modulated by drugs such as ciclopirox or deferiprone which might be repositioned to control cancer cell growth.