Comparative proteomics analysis of the antitumor effect of CIGB-552 peptide in HT-29 colon adenocarcinoma cells

• A pro-apoptotic effect of CIGB-552 peptide in colon cancer is proposed.
• An anti-inflammatory effect of CIGB-552 peptide in colon cancer is proposed.
• A relevance of NF-kB inhibition as mediator of CIGB-552 peptide effect in colon cancer is proposed.
• Application of DF-PAGE peptide fractionation method in the study of a novel drug action mechanism

The second generation peptide CIGB-552 has a pro-apoptotic effect on H460 non-small cell lung cancer cells and displays a potent cytotoxic effect in HT-29 colon adenocarcinoma cells though its action mechanism is ill defined. Here, we present the first proteomic study of peptide effect in HT-29 cells using subcellular fractionation, protein and peptide fractionation by DF-PAGE and LC–MS/MS peptide identification. In particular, we explored the nuclear proteome of HT-29 cells at a 5 h treatment identifying a total of 68 differentially modulated proteins, 49 of which localize to the nucleus. The differentially modulated proteins were analyzed following a system biology approach. Results pointed to a modulation of apoptosis, oxidative damage removal, NF-κB activation, inflammatory signaling and of cell adhesion and motility. Further Western blot and flow-cytometry experiments confirmed both pro-apoptotic and anti-inflammatory effects of CIGB-552 peptide in HT-29 cells.

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