The histone deacetylase inhibitor belinostat (PXD101) suppresses bladder cancer cell growth in vitro and in vivo

BackgroundTreatment options for patients with recurrent superficial bladder cancer are limited, necessitating aggressive exploration of new treatment strategies that effectively prevent recurrence and progression to invasive disease. We assessed the effects of belinostat (previously PXD101), a novel histone deacetylase inhibitor, on a panel of human bladder cancer cell lines representing superficial and invasive disease, and on a transgenic mouse model of superficial bladder cancer.MethodsGrowth inhibition and cell cycle distribution effect of belinostat on 5637, T24, J82, and RT4 urothelial lines were assessed. Ha-ras transgenic mice with established superficial bladder cancer were randomized to receive either belinostat or vehicle alone, and assessed for bladder weight, hematuria, gene expression profiling, and immunohistochemistry (IHC).ResultsBelinostat had a significant linear dose-dependent growth inhibition on all cell lines (IC50 range of 1.0–10.0 μM). The 5637 cell line, which was derived from a superficial papillary tumor, was the most sensitive to treatment. Belinostat (100 mg/kg, intraperitoneal, 5 days each week for 3 weeks) treated mice had less bladder weight (p < 0.05), and no hematuria compared with 6/10 control mice that developed at least one episode. IHC of bladder tumors showed less cell proliferation and a higher expression of p21WAF1 in the belinostat-treated mice. Gene expression profile analysis revealed 56 genes significantly different in the treated group; these included the upregulation of p21WAF1, induction of core histone deacetylase (HDAC), and cell communication genes.ConclusionOur data demonstrate that belinostat inhibits bladder cancer and supports the clinical evaluation of belinostat for the treatment of patients with superficial bladder cancer.

[1]  D. Budman,et al.  Review: Tubulin Function, Action of Antitubulin Drugs, and New Drug Development , 2005, Cancer investigation.

[2]  M. Sehested,et al.  Activity of PXD101, a histone deacetylase inhibitor, in preclinical ovarian cancer studies , 2006, Molecular Cancer Therapeutics.

[3]  J. Schaack,et al.  Suppression of adiponectin gene expression by histone deacetylase inhibitor valproic acid. , 2006, Endocrinology.

[4]  T. Sun,et al.  Role of Ha-ras activation in superficial papillary pathway of urothelial tumor formation , 2001, Oncogene.

[5]  Y. Ohshima,et al.  Histone deacetylase inhibitors such as sodium butyrate and trichostatin A induce apoptosis through an increase of the bcl-2-related protein Bad , 2006, Brain Tumor Pathology.

[6]  T. Sakai,et al.  Histone Deacetylase Inhibitor Activates the p21/WAF1/Cip1 Gene Promoter through the Sp1 Sites , 1999, Annals of the New York Academy of Sciences.

[7]  K. Glaser,et al.  Gene expression profiling of multiple histone deacetylase (HDAC) inhibitors: defining a common gene set produced by HDAC inhibition in T24 and MDA carcinoma cell lines. , 2003, Molecular cancer therapeutics.

[8]  Jun Luo,et al.  Hypermethylation and expression regulation of secreted frizzled-related protein genes in colorectal tumor. , 2006, World journal of gastroenterology.

[9]  R. Toillon,et al.  P21(WAF1/CIP1) is dispensable for G1 arrest, but indispensable for apoptosis induced by sodium butyrate in MCF-7 breast cancer cells. , 2004, Oncogene.

[10]  K. Ghoshal,et al.  Inhibitors of Histone Deacetylase and DNA Methyltransferase Synergistically Activate the Methylated Metallothionein I Promoter by Activating the Transcription Factor MTF-1 and Forming an Open Chromatin Structure , 2002, Molecular and Cellular Biology.

[11]  A. Kong,et al.  Identification of Nrf2-regulated genes induced by chemopreventive isothiocyanate PEITC by oligonucleotide microarray. , 2006, Life sciences.

[12]  E. Sausville,et al.  Phase 1 and pharmacologic study of MS-275, a histone deacetylase inhibitor, in adults with refractory and relapsed acute leukemias. , 2007, Blood.

[13]  T. Sun,et al.  Selective Interactions of UPIa and UPIb, Two Members of the Transmembrane 4 Superfamily, with Distinct Single Transmembrane-domained Proteins in Differentiated Urothelial Cells (*) , 1995, The Journal of Biological Chemistry.

[14]  P. Carroll,et al.  Evaluation of asymptomatic microscopic hematuria. , 1998, The Urologic clinics of North America.

[15]  K. Rieger,et al.  Identification of H-ras mutations in urine sediments complements cytology in the detection of bladder tumors. , 1995, Journal of the National Cancer Institute.

[16]  L. Schwartz,et al.  Phase I clinical trial of histone deacetylase inhibitor: suberoylanilide hydroxamic acid administered intravenously. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[17]  M. Sehested,et al.  The histone deacetylase inhibitor PXD101 synergises with 5-fluorouracil to inhibit colon cancer cell growth in vitro and in vivo , 2007, Cancer Chemotherapy and Pharmacology.

[18]  B. Czerniak,et al.  Concurrent mutations of coding and regulatory sequences of the Ha-ras gene in urinary bladder carcinomas. , 1992, Human pathology.

[19]  S. Formenti,et al.  Recombinant alpha2(IV)NC1 domain inhibits tumor cell-extracellular matrix interactions, induces cellular senescence, and inhibits tumor growth in vivo. , 2005, The American journal of pathology.

[20]  M. Mahadevappa,et al.  SARPs: a family of secreted apoptosis-related proteins. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  S. Bates,et al.  Phase I trial of the histone deacetylase inhibitor, depsipeptide (FR901228, NSC 630176), in patients with refractory neoplasms. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[22]  L. Ngo,et al.  Histone deacetylase (HDAC) inhibitor activation of p21WAF1 involves changes in promoter-associated proteins, including HDAC1. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[23]  L. Kiemeney,et al.  Gender differences in stage-adjusted bladder cancer survival. , 2000, Urology.

[24]  S. Formenti,et al.  Recombinant (cid:2) 2(IV)NC1 Domain Inhibits Tumor Cell-Extracellular Matrix Interactions, Induces Cellular Senescence, and Inhibits Tumor Growth in Vivo , 2005 .

[25]  P. Finn,et al.  Pharmacodynamic response and inhibition of growth of human tumor xenografts by the novel histone deacetylase inhibitor PXD101. , 2003, Molecular cancer therapeutics.

[26]  N. Socci,et al.  Molecular profiling of bladder cancer using cDNA microarrays: defining histogenesis and biological phenotypes. , 2002, Cancer research.

[27]  A. Kong,et al.  Toxicogenomics of endoplasmic reticulum stress inducer tunicamycin in the small intestine and liver of Nrf2 knockout and C57BL/6J mice. , 2007, Toxicology letters.

[28]  S. Ramalingam,et al.  Phase I and Pharmacokinetic Study of Vorinostat, A Histone Deacetylase Inhibitor, in Combination with Carboplatin and Paclitaxel for Advanced Solid Malignancies , 2007, Clinical Cancer Research.

[29]  P. Elliott,et al.  Proteasome inhibition with bortezomib (PS-341): a phase I study with pharmacodynamic end points using a day 1 and day 4 schedule in a 14-day cycle. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  O. Yokosuka,et al.  Cell Growth Inhibition and Gene Expression Induced by the Histone Deacetylase Inhibitor, Trichostatin A, on Human Hepatoma Cells , 2004, Oncology.

[31]  S. Lupold,et al.  Valproic acid, a histone deacetylase inhibitor, is an antagonist for oncolytic adenoviral gene therapy. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[32]  C. Cordon-Cardo,et al.  Hyperactivation of Ha-ras oncogene, but not Ink4a/Arf deficiency, triggers bladder tumorigenesis. , 2007, The Journal of clinical investigation.

[33]  C. Cordon-Cardo,et al.  Prostate cancer cell cycle regulators: response to androgen withdrawal and development of androgen independence. , 1999, Journal of the National Cancer Institute.

[34]  P. Marks,et al.  Histone deacetylases and cancer: causes and therapies , 2001, Nature Reviews Cancer.

[35]  Aimee M. Johnson,et al.  Early detection and measurement of urothelial tumors in mice. , 2006, Urology.

[36]  A. Jemal,et al.  Cancer Statistics, 2004 , 2004, CA: a cancer journal for clinicians.

[37]  Xue-Ru Wu.  Urothelial tumorigenesis: a tale of divergent pathways , 2005, Nature Reviews Cancer.

[38]  T. Griffith,et al.  Histone deacetylase inhibitors modulate the sensitivity of tumor necrosis factor-related apoptosis-inducing ligand-resistant bladder tumor cells. , 2006, Cancer research.

[39]  Y. Li,et al.  Valproic Acid Inhibits Invasiveness in Bladder Cancer but Not in Prostate Cancer Cells , 2006, Journal of Pharmacology and Experimental Therapeutics.

[40]  Hong-ying Huang,et al.  Uroplakin and androgen receptor expression in the human fetal genital tract: insights into the development of the vagina. , 2000, The Journal of urology.

[41]  Rune Matthiesen,et al.  Quantitative Proteomic Analysis of Post-translational Modifications of Human Histones* , 2006, Molecular & Cellular Proteomics.

[42]  T. Sun,et al.  Urothelium-specific expression of an oncogene in transgenic mice induced the formation of carcinoma in situ and invasive transitional cell carcinoma. , 1999, Cancer research.

[43]  Xu Zhang,et al.  Histone deacetylase inhibitor trichostatin A inhibits the growth of bladder cancer cells through induction of p21WAF1 and G1 cell cycle arrest , 2006, International journal of urology : official journal of the Japanese Urological Association.

[44]  P. Marks,et al.  Histone deacetylase inhibitors induce growth suppression and cell death in human rhabdomyosarcoma in vitro. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[45]  P. Marks,et al.  Histone deacetylase inhibitor selectively induces p21WAF1 expression and gene-associated histone acetylation. , 2000, Proceedings of the National Academy of Sciences of the United States of America.