Abstract

Maslinic acid (MA) is an anti-tumoural agent which shows potent anti-proliferative properties against the HT29 colon-cancer cells. To shed light upon the active mechanism of MA we have investigated its effects upon the cytoskeleton. We used a proteomics procedure based on two-dimensional gel electrophoresis, mass analysis and peptide mass fingerprinting. The incubation of HT29 cells with MA led to G1 cell-cycle arrest. After 24hours' exposure to 3.7μM (IC50/8) and 30μM (IC50) MA fourteen differentially expressed cytoskeletal proteins could be discerned. One group of these proteins, made up of keratin 2, keratin 8, keratin type II cytoskeletal 8, keratin type I cytoskeletal 9, keratin type I cytoskeletal 18, cytokeratins 18 and 19, and β-actin, exert a structural function, whilst another group, made up of lamin B1, gelsolin 1, septin 2, villin 1, actin-related protein 2 and moesin, is related to the nucleation of actin and cytoskeleton formation. Changes in the expression of moesin, villin 1 and β-actin identified by the proteomics techniques were corroborated by Western blotting. This is the first evidence obtained of the regulatory effects of MA on the cytoskeleton, which may prove to be one of the bases of its anti-proliferative effect against colon-cancer cells. In this paper we describe the changes in the expression of different cytoskeleton proteins identified by the proteomics techniques and corroborated by Western blotting. This is the first evidence obtained of the regulatory effects of MA on the cytoskeleton, which may prove to be one of the bases of its anti-proliferative effect against colon-cancer cells.

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