Investigation of the Effects of the Histone Deacetylase Inhibitor Saha on the Medulloblastoma Cell Line Daoy Using Gel-Based Proteomics
Suberoylanilide hydroxamic acid (SAHA) belongs to the class of histone deacetylase inhibitors emerging as potent antitumor agents. In this study, effects of SAHA on DAOY cells as model cell line for medulloblastoma, one of the most common malignant pediatric brain tumors, were analysed at the protein level by using gel-based proteomics. A 2D proteome map of DAOY cells in the pH range of 4-7 was created using 2D electrophoresis combined with MALDI-TOF/ TOF-MS. The 2D reference map of control and SAHA treated DAOY cells comprises 1196 identified protein spots of 778 distinct proteins. This is the first report of a 2D proteome map of SAHA treated DAOY cells and the largest catalogue of proteins identified in DAOY cells so far. Quantitative analysis of SAHA induced effects on the cells was performed using 2D-DIGE technique. After treatment with 10 μM SAHA, alterations were observed for 220 protein spots. Especially, SAHA induced fragmentation of many known caspase substrates. Higher levels of mitochondrial proteins indicated apoptosis via the mitochondrial pathway. Altogether, these findings point to induction of various antitumor activities by SAHA.
KeywordsDAOY gel-based proteomics medulloblastoma SAHA
Unable to display preview. Download preview PDF.