HDAC-inhibitor (S)-8 disrupts HDAC6-PP1 complex prompting A375 melanoma cell growth arrest and apoptosis.
Histone deacetylase inhibitors (HDACi) are agents capable of inducing growth arrest and apoptosis in different tumour cell types. Previously, we reported a series of novel HDACi obtained by hybridizing SAHA or oxamflatin with 1,4-benzodiazepines. Some of these hybrids proved effective against haematological and solid cancer cells and, above all, compound (S)-8 has emerged for its activities in various biological systems. Here, we describe the effectiveness of (S)-8 against highly metastatic human A375 melanoma cells by using normal PIG1 melanocytes as control. (S)-8 prompted: acetylation of histones H3/H4 and α-tubulin; G0 /G1 and G2 /M cell cycle arrest by rising p21 and hypophos-phorylated RB levels; apoptosis involving the cleavage of PARP and caspase 9, BAD protein augmentation and cytochrome c release; decrease in cell motility, invasiveness and pro-angiogenic potential as shown by results of wound-healing assay, down-regulation of MMP-2 and VEGF-A/VEGF-R2, besides TIMP-1/TIMP-2 up-regulation; and also intracellular accumulation of melanin and neutral lipids. The pan-caspase inhibitor Z-VAD-fmk, but not the antioxidant N-acetyl-cysteine, contrasted these events. Mechanistically, (S)-8 allows the disruption of cytoplasmic HDAC6-protein phosphatase 1 (PP1) complex in A375 cells thus releasing the active PP1 that dephosphorylates AKT and blocks its downstream pro-survival signalling. This view is consistent with results obtained by: inhibiting PP1 with Calyculin A; using PPP1R2-transfected cells with impaired PP1 activity; monitoring drug-induced HDAC6-PP1 complex re-shuffling; and, abrogating HDAC6 expression with specific siRNA. Altogether, (S)-8 proved very effective against melanoma A375 cells, but not normal melanocytes, and safe to normal mice thus offering attractive clinical prospects for treating this aggressive malignancy.
Department of Experimental and Clinical Medicine, University of Florence, Firenze, Italy.
Manjola
M
Balliu
Luca
L
Guandalini
Maria Novella
MN
Romanelli
Massimo
M
D'Amico
Francesco
F
Paoletti
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001570:Benzodiazepinones / Q000737:chemistry / Q000494:pharmacology*; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003984:Dibenzazepines / Q000737:chemistry / Q000494:pharmacology*; D005260:Female; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D008297:Male; D008544:Melanocytes / Q000187:drug effects / Q000473:pathology; D008545:Melanoma / Q000473:pathology*; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects; D054645:Protein Phosphatase 1 / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013237:Stereoisomerism
D001570:Benzodiazepinones; D003984:Dibenzazepines; D056572:Histone Deacetylase Inhibitors; C000600790:N1-hydroxy-N8-(1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo(e)(1,4)diazepin-3-yl)octanediamide; D054645:Protein Phosphatase 1; D020169:Caspases; C433811:HDAC6 protein, human; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
A375 human melanoma cells; AKT; HDAC-inhibitor (S)-8; HDAC6; HDAC6-PP1 complex; apoptosis; differentiation; growth arrest; in vivo toxicity; protein phosphatase 1 (PP1)