title: Proteomics in HepG2 hepatocarcinoma cells with stably silenced expression of PRDX1.
abstract: Peroxiredoxin 1 (PRDX1) is a member of the peroxiredoxin family. Aberrant expression of PRDX1 has been described in various cancers. We investigated the significance of this up-regulation in non-challenged hepatocellularcarcinoma (HCC) cells by establishing a HepG2 cell line stably expressing a Prdx1 shRNA. Prdx1 silencing reversed, at least partially, the tumoural phenotype of HepG2 cells, resulting in morphological changes, delayed cell growth, down-regulation of transcripts for AFP, osteopontin and β-catenin and decreased γ-glutamyl transpeptidase activity, and oppositely up-regulation of transcripts for E-cadherin and proapoptotic proteins (BAX, CASP3) and increased alkaline phosphatase and CASP3 activities. Proteomic profiling identified 16 spots differentially expressed in Prdx1-silenced cells. Most of the variations involved the down-regulation of proteins with pivotal roles in cell proliferation and differentiation, in agreement with the observed phenotypic changes. We also investigated the effect of Prdx1 silencing on thiol protein oxidation. Proteins prone to reversible cysteine oxidation play major physiological functions. Notably, the down-regulation and altered redox status of key enzymes of carbohydrate and amino acid metabolism suggested a disturbance of the Warburg effect and glutamine utilization, two major pathways in the proliferation of tumour cells. Overall, these observations suggest that PRDX1 acts as a pro-cancer protein in HCC HepG2 cells.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Córdoba, Campus de Rabanales, Edificio Severo Ochoa, planta 2(a)ª, Carretera Madrid-Cádiz Km 396-a, 14071-Córdoba, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Aguilar-Melero
authors.forename: María-José
authors.initials: MJ
authors.lastname: Prieto-Álamo
authors.forename: Juan
authors.initials: J
authors.lastname: Jurado
authors.forename: Arne
authors.initials: A
authors.lastname: Holmgren
authors.forename: Carmen
authors.initials: C
authors.lastname: Pueyo
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation; D015536:Down-Regulation; D056945:Hep G2 Cells / Q000378:metabolism*; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D054464:Peroxiredoxins / Q000235:genetics*; D040901:Proteomics; D034741:RNA, Small Interfering / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D034741:RNA, Small Interfering; C515940:PRDX1 protein, human; D054464:Peroxiredoxins