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Here, we compared the mode of action of romidepsin (FK228), an established class I histone deacetylase inhibitor, and domatinostat (4SC-202), a novel inhibitor of class I HDACs, which has been reported to also target the lysine-specific histone demethylase 1A (LSD1).

We performed MTS assays and flow cytometric analyses of propidium iodide or annexin V-stained cells to assess drug impact on cellular proliferation, cell cycle distribution, and survival.

While histone acetylation is generally associated with promoter activation, the role of another epigenetic modification, i.e., histone methylation, in regulation of gene expression is more complex and highly dependent on the specific histone residues modified [17].

In this regard, tri-methylation of lysine 9 or lysine 27 of histone H3 (H3K9me3 or H3K27me3) facilitates gene repression while di- or tri-methylation of lysine 4 of histone H3 (H3K4me2 or H3K4me3) mediates promoter activation, whereas mono-methylation of H3K4 is a hallmark of active distal enhancer regions [18].

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Gene expression analysis was performed using Nanos String technology.Targeting epigenetic modifiers is effective in cutaneous T cell lymphoma (CTCL).However, there is a need for further improvement of this therapeutic approach.Since time-lapse microscopy revealed that 4SC-202 could affect mitotic spindle formation, we performed an in vitro tubulin polymerization assay revealing that 4SC-202 can directly inhibit microtubule formation.We demonstrate that 4SC-202, a drug currently tested in clinical trials, effectively inhibits growth of CTCL cells.

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Recent genomic analysis revealed aberrations affecting apoptosis [6], cytokine, and T cell receptor signaling [7] as well as epigenetic regulation [8] as possible drivers in CTCL.

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