A proteomic analysis of differential cellular responses to the short-chain fatty acids butyrate, valerate and propionate in colon epithelial cancer cellsElectronic supplementary information (ESI) available: ESI 1: HCA data for cell-cycle, cytoskeletal fibre integrity and Keratin 8 fluorescence. ESI 2: Flow cytometry iTRAQ Workflow and correlation plots. ESI 3: Full list of proteins for iTRAQ Quant analysis. Phenyx v2.6 (GeneBio) is available via the internet at http://phenyx.vital-it.ch/pwi/logi

• Main Authors: Kilner, Josephine, Waby, Jennifer S, Chowdry, Joanna, Khan, Abdul Q, Noirel, Josselin, Wright, Phillip C, Corfe, Bernard M, Evans, Caroline A
• Format: Journal Article
• Language: English
• Published: 01.04.2012
• ISSN: 1742-206X; 1742-2051
• DOI: 10.1039/c1mb05219e

The short chain fatty acids (SCFAs) are inhibitors of histone deacetylases (HDACi); they are produced naturally in the colon by fermentation. They affect cellular processes at a molecular and transcriptional level, the mechanisms of which may involve large numbers of proteins and integrated pathways. Butyrate is the most biologically potent of the SCFAs in colon epithelial cells, inhibiting human colon carcinoma cell proliferation and inducing apoptosis in vitro . In order to investigate the hypothesis that propionate and valerate possess unique and independent actions from butyrate, we combined proteomic and cellomic approaches for large-scale comparative analysis. Proteomic evaluation was undertaken using an iTRAQ tandem mass-spectrometry workflow and high-throughput High-content Analysis microscopy (HCA) was applied to generate cellomic information on the cell cycle and the cytoskeletal structure. Our results show that these SCFAs possess specific effects. Butyrate was shown to have more pronounced effects on the keratins and intermediate filaments (IFs); while valerate altered the -tubulin isotypes' expression and the microtubules (MTs); propionate was involved in both mechanisms, displaying intermediate effects. These data suggest distinct physiological roles for SCFAs in colon epithelial function, offering new possibilities for cancer therapeutics. Integrated proteomic and cellomic workflows reveal that SCFA have separate and distinct effects on the cell cycle and cytoskeletons.