Proteomic analysis of pancreatic ductal carcinoma cells treated with 5-aza-2'-deoxycytidine

• Published in: Electrophoresis 2003-12, Vol.24 (24), p.4291-4303
• Main Authors: Cecconi, Daniela, Astner, Hubert, Donadelli, Massimo, Palmieri, Marta, Missiaglia, Edoardo, Hamdan, Mahmoud, Scarpa, Aldo, Righetti, Pier Giorgio
• Format: Article
• Language: English
• Subjects: Actin Depolymerizing Factors; Azacitidine - analogs & derivatives; Azacitidine - pharmacology; Carcinoma, Pancreatic Ductal - drug therapy; Carcinoma, Pancreatic Ductal - metabolism; Cell Division - drug effects; Contractile Proteins - metabolism; Cyclin-Dependent Kinase Inhibitor p16 - genetics; Cyclin-Dependent Kinase Inhibitor p16 - metabolism; Cyclophilin A - metabolism; Cystatin B; Cystatins - metabolism; DNA Methylation - drug effects; Ductal carcinomas; Humans; Mass Spectrometry; Microfilament Proteins - metabolism; Pancreatic Neoplasms - drug therapy; Pancreatic Neoplasms - metabolism; Pancreatic tumors; Profilins; Promoter Regions, Genetic - genetics; Proteomics; Rosaniline Dyes - chemistry; Superoxide Dismutase - metabolism; Tumor Cells, Cultured
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.200305724

A pancreatic adenocarcinoma cell line (PaCa44), which contains, among other alterations, a methylated p16 promoter, was treated with a chemoterapeutic agent, 5‐aza‐2'‐deoxycytidine (DAC), in order to evaluate the effect of this drug on cell growth and protein expression. Cell proliferation was strongly inhibited by a 24 h DAC treatment and this inhibition lasted for at least 10 days. Master maps of control and treated PaCa44 cells were generated by analysis with the PDQuest software. The comparison between such maps showed up‐ and downregulation of 45 polypeptide chains, of which 32 were downregulated and 13 upregulated, out of a total of 700 spots detected by a medium‐sensitivity stain, micellar Coomassie Brilliant Blue. Fingerprinting by mass spectrometry analysis enabled the identification of 36 of these spots. Among the major changes in DAC‐treated cells: cofilin and profilin 1 are silenced; coactosin, peptidyl‐propyl cis‐trans isomerase A and cystatin B are decreased by 22, 16‐ and 15‐fold, respectively; stress‐70 protein, superoxide dismutase and protein disulfide isomerase A3 are increased by 13‐, 11‐, and 5‐fold, respectively. The significance of some of these major changes is discussed.