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Since hydroxamate derivatives are also known as iron siderophores, it raises the possibility that WMJ-S-001 chelates iron which contributes to the activation of p53. Class I HDAC levels (HDAC1, 2, 3 and 8) are observed in various types of cancers and are associated with a poor prognosis.

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Immunoblotting analysis demonstrated that p21(A) Cells were treated with vehicle or WMJ-S-001 at 10 μM for various time periods as indicated. Compiled results are shown at the bottom of the chart. The compiled results of AMPK and p38MAPK phosphorylations are shown. p53 stability and activity are dependent on post-transcriptional modifications such as phosphorylation, sumoylation, ubiquitination and acetylation.

Longer exposure to WMJ-S-001 (48 h) further decreased HCT116 cell viability (Fig. In contrast, treatment of cells with WMJ-S-002, WMJ-S-003, WMJ-S-004, or WMJ-S-005 for 24 h only slightly affected cell viability (Fig. We sought to further investigate the mechanisms of HCT116 cell death after exposure to WMJ-S-001 in the following experiments.

To determine whether decreased HCT116 cell viability in the presence of WMJ-S-001 was a result of cell apoptosis, flow cytometric analysis with propidium iodide (PI) labeling was used. 1c, WMJ-S-001, at concentrations higher than 10 μM (20 and 30 μM), significantly increased the percentage of PI-stained cells in the apoptotic region (Apo, sub-G1/G1 peak). The full-length blot is presented in Supplementary Figure 5a–c (*p . 7a, WMJ-S-001 significantly inhibited serum-induced proliferation in Colo205 cells. 7b) levels were also reduced in HT29 cells as compared with Colo205 cells.

Transfection with AMPK dominant negative mutant (DN) reduced WMJ-S-001’s effects on p53 and Sp1 binding to the survivn promoter region.

Transfection with HDAC3-Flag or HDAC4-Flag also abrogated WMJ-S-001’s enhancing effect on p53 acetylation.

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