title: Belinostat, a potent HDACi, exerts antileukaemic effect in human acute promyelocytic leukaemia cells via chromatin remodelling.
abstract: Epigenetic changes play a significant role in leukaemia pathogenesis, therefore histone deacetylases (HDACis) are widely accepted as an attractive strategy for acute promyelocytic leukaemia (APL) treatment. Belinostat (Bel, PXD101), a hydroxamate-type HDACi, has proved to be a promising cure in clinical trials for solid tumours and haematological malignancies. However, insight into molecular effects of Bel on APL, is still lacking. In this study, we investigated the effect of Bel alone and in combination with differentiation inducer retinoic acid (RA) on human promyelocytic leukaemia NB4 and HL-60 cells. We found that treatment with Bel, depending on the dosage used, inhibits cell proliferation, whereas in combination with RA enhances and accelerates granulocytic leukaemia cell differentiation. We also evaluated the effect of used treatments with Bel and RA on certain epigenetic modifiers (HDAC1, HDAC2, PCAF) as well as cell cycle regulators (p27) gene expression and protein level modulation. We showed that Bel in combination with RA up-regulates basal histone H4 hyperacetylation level more strongly compared to Bel or RA alone. Furthermore, chromatin immunoprecipitation assay indicated that Bel induces the accumulation of hyperacetylated histone H4 at the p27 promoter region. Mass spectrometry analysis revealed that in control NB4 cells, hyperacetylated histone H4 is mainly found in association with proteins involved in DNA replication and transcription, whereas after Bel treatment it is found with proteins implicated in pro-apoptotic processes, in defence against oxidative stress and tumour suppression. Summarizing, our study provides some novel insights into the molecular mechanisms of HDACi Bel action on APL cells.
authors: Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Giedre
authors: G
authors: Valiuliene
authors: Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Ieva
authors: I
authors: Stirblyte
authors: Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Dovile
authors: D
authors: Cicenaite
authors: Proteomics Centre, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Algirdas
authors: A
authors: Kaupinis
authors: Proteomics Centre, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Mindaugas
authors: M
authors: Valius
authors: Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors: Ruta
authors: R
authors: Navakauskiene
mesh_terms: D000107:Acetylation / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006098:Granulocytes / Q000187:drug effects / Q000473:pathology; D018922:HL-60 Cells; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics / Q000473:pathology*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D013449:Sulfonamides / Q000494:pharmacology*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D009363:Neoplasm Proteins; D013449:Sulfonamides; D050760:Cyclin-Dependent Kinase Inhibitor p27; D014212:Tretinoin; C487081:belinostat
keywords: APL; HDACi; belinostat; epigenetics; granulocytic differentiation