Figure 1
Induction of the CAMP gene by Entinostat in the Campluc reporter cells (a) and in HT-29 cells (b,c). (a) CampLuc reporter cells were stimulated for 24 h with the HDAC inhibitors phenylbutyrate (PBA, 2 mM), isovaleric acid (50 μM–1 mM), isobutyric acid (100 μM–2 mM), vorinostat (Vorino, 5–250 μM), Entinostat (Entino, 250–2.5 μM, black bars), and valproic acid (VPA, 0.5–2 mM). Vitamin D3 (Vit D, 100 nM) was also tested, alone or in combination with PBA (2 mM) or Entinostat (2.5 μM, black bar). The luciferase activity was assayed, and the results are expressed as luminescence relative to vehicle (ctrl). Graph is representative of at least 3 experiments (Compiled with data from Nylen et al.19). (b) Stimulation of HT-29 cells by Entinostat (250 nM-1 mM), PBA (2 mM) and PBA/Entinostat in combination with Vitamin D3 (Vit D, 100 nM) at different concentrations was determined for induction of the CAMP gene expression by qRT-PCR. Significantly altered expression is indicated vs ctrl: *p < 0.05; induction vs PBA: #p < 0.05; induction vs Entinostat (Entino): ¤p < 0.05. (c) Treatment of HT-29 cells with Entinostat (2.5 μM) at different time-points (6–48 h) was analysed for the induction of CAMP gene expression by qRT-PCR. Significant altered expression vs ctrl is indicated: *p < 0.05. (d) The expression, of the genes DEFB1 and DEFB4 encoding HBD1 and HBD2, was assessed in HT-29 parental cells by qRT-PCR after stimulation with Entinostat (2.5 μM) or PBA (2 mM). Significantly altered expression vs ctrl is indicated: *p < 0.05.
