Salicylate, diflunisal and their metabolites inhibit CBP/p300 and exhibit anticancer activity
- University of California, United States
- Kyoto University, Japan
- University of Miami, United States
- Sylvester Comprehensive Cancer Center, United States
- University of Pennsylvania, United States
- Abramson Family Cancer Research Institute, United States
- National Cancer Institute, United States
Figures
Figure 1 with 1 supplement
Salicylate inhibits CBP/p300 in vitro.
(A) Recombinant p300, CBP, GCN5, or PCAF and histones were incubated with 14C-labeled acetyl-CoA with or without sodium salicylate, separated by SDS-PAGE, analyzed by autoradiography, and quantified …
Figure 1—figure supplement 1
CoA metabolites of salicylate and diflunisal are more potent inhibitors of p300.
(A) Dose-response data for inhibition of p300 by salicylate and salicyl-CoA. Acetylation of an H4(3–14) peptide was monitored using direct microfluidic mobility shift analysis as previously …
Figure 2 with 2 supplements
Salicylate inhibits specific lysine acetylation of histone and nonhistone proteins independently of AMPK activation.
(A) Decreased acetylation of specific lysines in histones in the presence of salicylate. HEK293T cells were treated with the indicated concentrations of sodium salicylate for 24 hr. Site-specific …
Figure 2—figure supplement 1
Salicylate induces histone deacetylation in HEK293T cells HEK293T cells were treated with sodium salicylate as indicated for 24 hr, immediately lysed in Laemmli buffer, and then subjected to western blot analysis with the indicated antibodies.
Histones H2A, H2B, H3 and H4 were used as input loading controls. Experiments are repeated five times and representative data are shown.
Figure 2—figure supplement 2
Salicylate-induced deacetylation of histone H2B can be rescued by overexpression of p300, but not PCAF, in a dose-dependent manner.
(A) Overexpression of p300 but not PCAF specifically leads to hyperacetylation of histone H2B. Expression plasmids for p300 WT, catalytically inactive (Y1503A or F1504A) p300, or PCAF were …
Figure 3 with 2 supplements
Structural homology search identifies diflunisal as a potent p300 inhibitor.
(A) FDA-approved drugs that contain a structure similar to that of salicylate are shown in red. Numbers below the structures are IC50 of each drug, measured by in vitro p300 HAT assays. (B) Relative …
Figure 3—figure supplement 1
Diflunisal induces histone deacetylation in HEK293T cells HEK293T cells were treated with diflunisal as indicated for 24 hr, immediately lysed in Laemmli buffer, and then subjected to western blot analysis with the indicated antibodies.
Histones H2A, H2B, H3 and H4 were used as input loading controls. Experiments are repeated five times and representative data are shown.
Figure 3—figure supplement 2
Diflunisal-induced deacetylation of p300 is rescued by overexpression of p300 in a dose-dependent manner, but not inactive p300 mutants.
HEK293T cells were transfected with expression vectors for WT p300 or catalitycally inactive mutant. 24 hr after transfection, cells were treated with diflunisal as indicated for 24 hr. H2B …
Figure 4
Sodium salicylate and diflunisal decrease acetylation of AML1-ETOK43/K24 and block the growth of t(8;21) leukemia cells by inducing apoptosis.
(A) Kasumi-1 cells expressing the AML1-ETO fusion protein were treated with sodium salicylate (4 or 8 mM, left) or diflunisal (100 or 200 μM, right) for 24 hr, followed by immunoprecipitation of …
Figure 5
Sodium salicylate and diflunisal inhibit the growth of AML1-ETO leukemia cells in SCID mice.
The mice were inoculated with Kasumi-1 cells (3x107) and, starting 3 weeks later, were treated daily with oral diflunisal (50 or 100 mg/kg) or vehicle for 3 weeks. (A) and (B) Plots of tumor volume …
