Belinostat

Manufactured by Selleck Chemicals

Sourced in United States

Belinostat is a chemical compound used in laboratory research. It is a histone deacetylase (HDAC) inhibitor, which means it can regulate gene expression by modifying the structure of chromatin. Belinostat is commonly used in cell-based and biochemical assays to study the effects of HDAC inhibition on various cellular processes.

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Belinostat (PXD101) (6 citations)

18 protocols using belinostat

Belinostat and Retinoic Acid in APL Cells

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The human APL cells NB4 and HL-60 (from DSMZ, GmbH, Braunschweig, Germany) were maintained in RPMI 1640 medium supplemented with 10% foetal bovine serum, 100 U/ml penicillin and 100 mg/ml streptomycin (Gibco, Grand Island, NY, USA) in a humidified incubator at 37°C with 5% CO₂. For each experiment, logarithmically growing cells were seeded at a density of 0.5 × 10⁶ cells/ml in 5 ml of medium. According to previous publication 22 cells were exposed to 0.2 and 2.0 μM Belinostat (Selleck Chemicals, Houston, TX, USA) alone or in combination with 1 μM RA (Sigma-Aldrich, St. Louis, MO, USA). The agents were left in the cell media for the duration of the experiment.

Valiuliene G., Stirblyte I., Cicenaite D., Kaupinis A., Valius M, & Navakauskiene R. (2015). Belinostat, a potent HDACi, exerts antileukaemic effect in human acute promyelocytic leukaemia cells via chromatin remodelling. Journal of Cellular and Molecular Medicine, 19(7), 1742-1755.

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Quantification of SN-38 and Metabolites

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SN-38, SN-38G and their internal standards, SN-38-d₃ and SN-38G-d₃ respectively, were obtained from Toronto Research Chemicals (Toronto, ON, Canada). Belinostat, vorinostat and panobinostat, were purchased from Selleck Chemicals (Houston, TX, USA). Human recombinant UDP-glucuronosyltransferases (UGT1A1, UGT1A6, UGT1A7, UGT1A9), UGT control, UGT reaction mix - solutions A and B, HLMs (50 donor pool, UGT1A1*1*28 and UGT1A1*28*28 allelic variants) were purchased from BD Gentest (San Jose, CA, USA). All other reagents were of HPLC grade or of the highest grade commercially available.

Wang L., Linus Chan C.E., Wong A.L., Wong F.C., Lim S.W., Chinnathambi A., Alharbi S.A., Lee L.S., Soo R., Yong W.P., Lee S.C., Ho P.C., Sethi G, & Goh B.C. (2017). Combined use of irinotecan with histone deacetylase inhibitor belinostat could cause severe toxicity by inhibiting SN-38 glucuronidation via UGT1A1. Oncotarget, 8(25), 41572-41581.

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Dissolution and Storage Protocols

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Vorinostat and fluvastatin purchased from Cayman Chemical, panobinostat purchased from LC Laboratories, belinostat purchased from Selleck Chemicals, and tunicamycin purchased from Enzo Life Sciences were dissolved in DMSO. Compound C dihydrochloride purchased from R&D Systems and cycloheximide purchased from Enzo Life Sciences were dissolved in distilled water. These reagents were stored at −80°C or −20°C until use.

Okubo K., Isono M., Miyai K., Asano T, & Sato A. (2019). Fluvastatin potentiates anticancer activity of vorinostat in renal cancer cells. Cancer Science, 111(1), 112-126.

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Establishment and Characterization of Testicular Cancer Cell Lines

Cited 2 times

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The (T)GCT cell lines (n = 4) TCam-2 (a SE-like cell line) and NCCIT, 2102Ep, and NT2 (representative of NS) were kindly provided by Prof. Leendert Looijenga. These cell lines have been previously characterized, including copy number alterations, and were cultured as described [66 (link)]. Additionally, an independent set of matched clones of cisplatin-sensitive and cisplatin-resistant cell lines (n = 6) were kindly provided by Prof. Daniel Nettersheim and generated by Dr. Christoph Oing and Prof. Friedemann Honecker. The resistant clones (NCCIT-R, 2102Ep-R, and NT2-R) were obtained from the parental matched clone (NCCIT-P, 2102Ep-P, and NT2-P) upon culturing with increasing doses of cisplatin, as reported before [67 (link)]. Cells were maintained in low passages and were negative for Mycoplasma spp. (Clontech Laboratories; Mountain View, CA, USA; tested twice a month). Cisplatin was kindly provided by IPO Porto’s Department of Pharmacy. Belinostat and panobinostat were purchased from Selleckchem, Houston, TX, USA (Catalog No. S1085 and Catalog No. S1030, respectively).

Lobo J., Guimarães-Teixeira C., Barros-Silva D., Miranda-Gonçalves V., Camilo V., Guimarães R., Cantante M., Braga I., Maurício J., Oing C., Honecker F., Nettersheim D., Looijenga L.H., Henrique R, & Jerónimo C. (2020). Efficacy of HDAC Inhibitors Belinostat and Panobinostat against Cisplatin-Sensitive and Cisplatin-Resistant Testicular Germ Cell Tumors. Cancers, 12(10), 2903.

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Quantitative Drug Response Assay

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Tariquidar, paclitaxel, doxorubicin, orlistat, JQ1, AS1842856, linsitinib, panobinostat, vorinostat, belinostat, and LMK-235 (all from Selleckchem, Houston, TX, USA) and tepoxalin (Toronto Research Chemicals, North York, ON, CA) were prepared in DMSO. Metformin and 2-deoxy-D-glucose (both from Selleckchem) were prepared in complete culture medium. Prior to plating for drug response assays, breast cancer cell lines transduced with tet-inducible eGFP or ABCB1 lentiviruses were exposed to 1 µg/mL doxycycline for 24 hours to induce gene expression. After 24 hours of doxycycline exposure, 5,000 cells/well were seeded in 30 µL of RPMI + 10% FBS in 384-well microplates (Corning cat. no. 3764, Corning, NY, USA). After 24 hours, 10 µL of 4X drug was added in quadruplicate per dose. Upon 72 hours of drug treatment, 30 µL CellTiterGlo (Promega, Madison, WI, USA) was added and luminescence assessed according to manufacturer's directions using an Infinite M1000 Pro plate reader (Tecan, Morrisville, NC, USA).

McQuerry J.A., Chen J., Chang J.T, & Bild A.H. (2021). Tepoxalin increases chemotherapy efficacy in drug-resistant breast cancer cells overexpressing the multidrug transporter gene ABCB1. Translational Oncology, 14(10), 101181.

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Protein Modulator Screening Assay

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Twenty-four hours after seeding, cells were treated with BTZ (Selleckchem), CFZ (Selleckchem), MG132 (Selleckchem), givinostat (Selleckchem), belinostat (Selleckchem), tubacin (Selleckchem), Lumacaftor (VX-809; Selleckchem), Ivacaftor (VX-770; Selleckchem) or the carrier 0.1% dimethyl sulfoxide (DMSO, VWR). Cells were analyzed after 24 h of treatment.

Hoch L., Bourg N., Degrugillier F., Bruge C., Benabides M., Pellier E., Tournois J., Mahé G., Maignan N., Dawe J., Georges M., Papazian D., Subramanian N., Simon S., Fanen P., Delevoye C., Richard I, & Nissan X. (2022). Dual Blockade of Misfolded Alpha-Sarcoglycan Degradation by Bortezomib and Givinostat Combination. Frontiers in Pharmacology, 13, 856804.

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HDAC Inhibitors and Chloroquine Synergy

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Vorinostat (SAHA) and chloroquine diphosphate salt were purchased from Sigma-Aldrich (USA). Romidepsin (FK228), Belinostat (PXD101), and Panobinostat (LBH589) were purchased from Selleck Chemicals. All HDAC inhibitors were prepared as 10–20 mM stock solutions in 100% DMSO. Chloroquine was prepared as a 10–20 mM stock in phosphate buffered saline (PBS).

Chua M.J., Arnold M.S., Xu W., Lancelot J., Lamotte S., Späth G.F., Prina E., Pierce R.J., Fairlie D.P., Skinner-Adams T.S, & Andrews K.T. (2016). Effect of clinically approved HDAC inhibitors on Plasmodium, Leishmania and Schistosoma parasite growth. International Journal for Parasitology: Drugs and Drug Resistance, 7(1), 42-50.

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Inhibitor Screening for Epigenetic Targets

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Chidamide was provided by Shenzhen Chipscreen Biosciences Ltd. SAHA was obtained from Sigma–Aldrich. Trichostatin A (TSA), belinostat and tofacitinib were purchased from Selleckchem. Romidepsin was obtained from MedChemExpress. Ruxolitinib and Stattic were purchased from Axon Medchem and Millipore, respectively.

Chen J., Zuo Z., Gao Y., Yao X., Guan P., Wang Y., Li Z., Liu Z., Hong J.H., Deng P., Chan J.Y., Cheah D.M., Lim J., Chai K.X., Chia B.K., Pang J.W., Koh J., Huang D., He H., Sun Y., Liu L., Liu S., Huang Y., Wang X., You H., Saraf S.A., Grigoropoulos N.F., Li X., Bei J., Kang T., Lim S.T., Teh B.T., Huang H., Ong C.K, & Tan J. (2023). Aberrant JAK-STAT signaling-mediated chromatin remodeling impairs the sensitivity of NK/T-cell lymphoma to chidamide. Clinical Epigenetics, 15, 19.

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Chemical Compounds for Cancer Research

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Sodium-butyrate and valproate were obtained from Sigma (St. Louis, MO). Vorinostat, belinostat, Depsipeptide, entinostat, ABT-737 and ABT-199 were obtained from Selleck Chemicals (Houston, TX). ABT-263 was obtained from ApexBio (USA). Synthesis of A-1331852 was as described previously (26 ).

Chueh A.C., Tse J.W., Dickinson M., Ioannidis P., Jenkins L., Togel L., Tan B., Luk I., Davalos-Salas M., Nightingale R., Thompson M.R., Williams B.R., Lessene G., Lee E., Fairlie W.D., Dhillon A.S, & Mariadason J.M. (2017). ATF3 repression of BCL-XL determines apoptotic sensitivity to HDAC inhibitors across tumour types. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(18), 5573-5584.

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Evaluating HDAC Inhibitor Effects on Ototoxicity

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Explants were incubated for 2 days to recover from dissection stress before administering HDAC inhibitors and gentamicin (GM; Sigma-Aldrich Co., St. Louis, MO, USA). The medium was then exchanged for new medium containing a final concentration of 4.5 μM GM with or without various concentrations of the HDAC inhibitors and incubated for 72 h. The stock solutions with 95 mM of SAHA/vorinostat (Cayman Chemical, Ann Arbor, MI, USA), 100 mM of belinostat (Selleckchem, Houson, TX, USA), and 100 mM of panobinostat (Cayman Chemical), were dissolved in 100% dimethyl sulfoxide (DMSO) and stored at −20°C. GM solution was made fresh from powder in culture medium at 0.2 mM and then diluted to the final concentration for each experiment.

Yang C.H., Liu Z., Dong D., Schacht J., Arya D, & Sha S.H. (2017). Histone Deacetylase Inhibitors Are Protective in Acute but Not in Chronic Models of Ototoxicity. Frontiers in Cellular Neuroscience, 11, 315.

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