title: A combined free-flow electrophoresis and DIGE approach to identify proteins regulated by butyrate in HT29 cells.
abstract: Many biologically active agents exert a pleiotropic response in cells and tissues. This presents challenges in descriptive and comparative analysis of the proteome in response to these agents. Although free-flow electrophoresis has been applied in a number of proteomic studies as a protein separation technique, the combination of free-flow electrophoresis and DIGE has not yet been investigated for comparative proteomic analysis. In this study, we have compared the effects of butyrate on HT29 colorectal cancer cells with a particular focus on apoptosis and describe the utility of a novel approach combining free-flow electrophoresis with DIGE to identify differentially expressed proteins. We verify the results obtained by the combined free-flow electrophoresis and DIGE approach with Western blot analysis of selected proteins. We also report for the first time the regulation of a number of proteins by butyrate in HT29 colorectal cells including peptidyl-prolyl cis-trans isomerase A (cyclophilin A) and profilin-1.
authors.affiliation: CSIRO, Preventative Health Flagship, Australia. kim.fung@csiro.au
authors.forename: Kim Y C
authors.initials: KY
authors.lastname: Fung
authors.forename: Chris
authors.initials: C
authors.lastname: Cursaro
authors.forename: Tanya
authors.initials: T
authors.lastname: Lewanowitsch
authors.forename: Gemma V
authors.initials: GV
authors.lastname: Brierley
authors.forename: Shaun R
authors.initials: SR
authors.lastname: McColl
authors.forename: Trevor
authors.initials: T
authors.lastname: Lockett
authors.forename: Richard
authors.initials: R
authors.lastname: Head
authors.forename: Peter
authors.initials: P
authors.lastname: Hoffmann
authors.forename: Leah
authors.initials: L
authors.lastname: Cosgrove
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002087:Butyrates / Q000494:pharmacology*; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control; D021984:Cyclophilin A / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D019073:HT29 Cells; D006801:Humans; D051304:Profilins / Q000235:genetics* / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology*; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D002087:Butyrates; D051304:Profilins; D020011:Protective Agents; D020543:Proteome; D021984:Cyclophilin A