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Histone deacetylase inhibitor potentiates chemotherapy-induced apoptosis through Bim upregulation in Burkitt’s lymphoma cells

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Abstract

Purpose

Although polychemotherapy regiments have improved clinical outcome for Burkitt’s lymphoma (BL) patients, salvage treatment of patients with refractory disease remains very poor. Combined therapies protocols have been emerging to improve treatment strategies to circumvent responseless BL patients. We evaluate the cell death effect of histone deacetylase inhibitor (HDACI) combined with etoposide (VP-16) and cisplatin (CDDP) on BL cell lines.

Methods

3-(4,5-Dimethyl-thiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay was performed to assess drug toxicity. To establish the concentrations and time of incubation for the combined treatment, a kinetic analysis was performed for each drug on BL41 and Raji BL cell lines for 24, 48 and 72 h. Apoptosis was assessed by flow cytometry using Annexin V/propidium iodide (PI) and cleaved caspase 3 labeling assays. Caspase 9 activation and levels of Bcl-2 family proteins were analyzed by Western blot.

Results

The doses of NaB (1.0 mM), CDDP (1.0 and 2.5 μM), and VP-16 (0.1 and 0.3 μM) after 24 h of incubation were chosen for the evaluation of combined therapy. The apoptotic effects on BL cell lines of NaB/VP-16 and NaB/CDDP were followed by upregulation of Bim protein (P < 0.05), activation of caspase-3 and caspase-9, followed by Mcl-1 downregulation (P < 0.05). However, Bim overexpression was not correlated with Bcl-2 inhibition (P > 0.05) and was accompanied by increase in Bax expression (P < 0.05). The combination effects of NaB/VP-16 and NaB/CDDP were found to be synergistic and additive, respectively, in both the cell lines.

Conclusions

The study provides strong evidence for the synergistic effects of the association with HDCI and chemotherapy in BL cells.

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References

  • Aldoss IT, Weisenburger DD, Fu K, Chan WC, Vose JM, Bierman PJ, Bociek RG, Armitage JO (2008) Adult Burkitt lymphoma: advances in diagnosis and treatment. Oncology 22:1508–1517

    PubMed  Google Scholar 

  • Atra A, Gerrard M, Hobson R, Imeson JD, Hann IM, Pinkerton CR (2001) Outcome of relapsed or refractory childhood B-cell acute lymphoblastic leukaemia and B-cell non-Hodgkin’s lymphoma treated with the UKCCSG 9003/9002 protocols. Br J Haematol 112:965–968. doi:10.1046/j.1365-2141.2001.02647.x

    Article  PubMed  CAS  Google Scholar 

  • Belloc F, Belaud-Rotureau MA, Lavignolle V, Bascans E, Braz-Pereira E, Durrieu F, Lacombe F (2000) Flow cytometry detection of caspase 3 activation in preapoptotic leukemic cells. Cytometry 40:150–151. doi:10.1002/(SICI)1097-0320(20000601)40:2<151:AID-CYTO9>3.0.CO;2-9

    Article  Google Scholar 

  • Bornkamm GW (2009) Epstein-Barr virus and its role in the pathogenesis of Burkitt’s lymphoma: an unresolved issue. Semin Cancer Biol 19:351–365. doi:10.1016/j.semcancer.2009.07.002

    Article  PubMed  CAS  Google Scholar 

  • Burkitt D (1958) A sarcoma involving the jaws in African children. Br J Surg 46:218–223

    Article  PubMed  CAS  Google Scholar 

  • Cairo MS, Sposto R, Perkins SL, Meadows AT, Hoover-Regan ML, Anderson JR, Siegel SE, Lones MA, Tedeschi-Blok N, Kadin ME, Kjeldsberg CR, Wilson JF, Sanger W, Morris E, Krailo MD, Finlay JL (2003) Burkitt’s and Burkitt-like lymphoma in children and adolescents: a review of the children’s cancer group experience. Br J Haematol 120:660–670

    Article  PubMed  Google Scholar 

  • Calender A, Billaud M, Aubry JP, Banchereau J, Vuillaume M, Lenoir GM (1987) Epstein-Barr virus (EBV) induces expression of B-cell activation markers on in vitro infection of EBV-negative B-lymphoma cells. Proc Natl Acad Sci USA 84:8060–8064

    Article  PubMed  CAS  Google Scholar 

  • Cang S, Ma Y, Liu D (2009) New clinical developments in histone deacetylase inhibitors for epigenetic therapy of cancer. J Hematol Oncol 2:22–32. doi:10.1186/1756-8722-2-22

    Article  PubMed  Google Scholar 

  • Carew JS, Giles FJ, Nawrocki ST (2008) Histone deacetylase inhibitors: mechanisms of cell death and promise in combination cancer therapy. Cancer Lett 269:7–17. doi:10.1016/j.canlet.2008.03.037

    Article  PubMed  CAS  Google Scholar 

  • Chen X, Wong P, Radany E, Wong JY (2009) HDAC Inhibitor, Valproic acid, induces p53-dependent radiosensitization of colon cancer cells. Cancer Biother Radiopharm 24:689–699. doi:10.1089/cbr.2009.0629

    Article  PubMed  CAS  Google Scholar 

  • Cherney BW, Bhatia KG, Sgadari C, Gutierrez MI, Mostowski H, Pike SE, Gupta G, Magrath IT, Tosato G (1997) Role of the p53 tumor suppressor gene in the tumorigenicity of Burkitt’s lymphoma cells. Cancer Res 57:2508–2515

    PubMed  CAS  Google Scholar 

  • Chou T, Talalay P (1984) Quantitative analysis of dose-effects relationships: the combined effects of multiple drugs or enzyme inhibtors. Adv Enzyme Regul 22:27–55

    Article  PubMed  CAS  Google Scholar 

  • Craig RW (2002) MCL1 provides a window on the role of the BCL2 family in cell proliferation, differentiation and tumorigenesis. Leukemia 16:444–454. doi:10.1038/sj/leu/2402416

    Article  PubMed  CAS  Google Scholar 

  • Czabotar PE, Colman PM, Huang DC (2009) Bax activation by Bim? Cell Death Differ 16:1187–1191. doi:10.1038/cdd.2009.83

    Article  PubMed  CAS  Google Scholar 

  • Ellis L, Pili R (2010) Histone deacetylase inhibitors: advancing therapeutic strategies in hematological and solid malignancies. Pharmaceuticals (Basel) 3:2411–2469. doi:10.3390/ph3082441

    Google Scholar 

  • Ellis L, Bots M, Lindemann RK, Bolden JE, Newbold A, Cluse LA, Scott CL, Strasser A, Atadja P, Lowe SW, Johnstone RW (2009) The histone deacetylase inhibitors LAQ824 and LBH589 do not require death receptor signaling or a functional apoptosome to mediate tumor cell death or therapeutic efficacy. Blood 114:380–393. doi:10.1182/blood-2008-10-182758

    Article  PubMed  CAS  Google Scholar 

  • Emanuele S, Lauricella M, Tesoriere G (2008) Histone deacetylase inhibitors: apoptotic effects and clinical implications Int J Oncol 33:637–646. doi:10.3892/ijo_00000049

    CAS  Google Scholar 

  • Fischel J-L, Formento P, Milano G (2005) Epidermal growth factorreceptor double targeting by a tyrosine kinase inhibitor (Iressa) and a monoclonal antibody (Cetuximab). Impact on cell growth and molecular factors Br J Cancer 92:1063–1068. doi:10.1038/sj.bjc.6602428

    CAS  Google Scholar 

  • Gerrard M, Cairo MS, Weston C, Auperin A, Pinkerton R, Lambilliote A, Sposto R, McCarthy K, Lacombe MJ, Perkins SL, Patte C (2008) FAB LMB96 International Study Committee. Excellent survival following two courses of COPAD chemotherapy in children and adolescents with resected localized B-cell non-Hodgkin’s lymphoma: results of the FAB/LMB 96 international study. Br J Haematol 141:840–847. doi:10.1111/j.1365-2141.2008.07144.x

    Google Scholar 

  • Griffin TC, Weitzman S, Weinstein H, Chang M, Cairo M, Hutchison R, Shiramizu B, Wiley J, Woods D, Barnich M, Gross TG, Group Children’sOncology (2009) A study of rituximab and ifosfamide, carboplatin, and etoposide chemotherapy in children with recurrent/refractory B-cell (CD20+) non-Hodgkin lymphoma and mature B-cell acute lymphoblastic leukemia: a report from the Children’s Oncology Group. Pediatr Blood Cancer 52:177–181. doi:10.1002/pbc.21753

    Article  PubMed  Google Scholar 

  • Guo F, Sigua C, Tao J, Bali P, George P, Li Y, Wittmann S, Moscinski L, Atadja P, Bhalla K (2004) Cotreatment with histone deacetylase inhibitor LAQ824 enhances Apo-2L/tumor necrosis factor-related apoptosis inducing ligand-induced death inducing signaling complex activity and apoptosis of human acute leukemia cells. Cancer Res 64:2580–2589. doi:10.1158/0008-5472

    Article  PubMed  CAS  Google Scholar 

  • Jazirehi AR (2010) Regulation of apoptosis-associated genes by histone deacetylase inhibitors: implications in cancer therapy. Anticancer Drugs 21:805–813. doi:10.1097/CAD.0b013e32833dad91

    Article  PubMed  CAS  Google Scholar 

  • Kelly GL, Rickinson AB (2007) Burkitt lymphoma: revisiting the pathogenesis of a virus-associated malignancy. Hematology 20:277–284, http://asheducationbook. hematologylibrary.org

    Google Scholar 

  • Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR, Newmeyer DD (2005) BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Molecular Cell 17:525–535. doi:10.1016/j.molcel.2005.02.003

    Article  PubMed  CAS  Google Scholar 

  • Lemoine M, Younes A (2010) Histone deacetylase inhibitors in the treatment of lymphoma. Discov Med 10:462–470

    PubMed  Google Scholar 

  • Magrath I, Jain V, Bhatia K (1982) Epstein-Barr virus and Burkitt’s lymphoma. Semin Cancer Biol 3:285–295

    Google Scholar 

  • Martínez-Iglesias O, Ruiz-Llorente L, Sánchez-Martínez R, García L, Zambrano A, Aranda A (2008) Histone deacetylase inhibitors: mechanism of action and therapeutic use in cancer. Clin Transl Oncol 10:395–398

    Article  PubMed  Google Scholar 

  • Mellert HS, Stanek TJ, Sykes SM, Rauscher FJ, Schultz DC, McMahon SB (2011) Deacetylation of the DNA-binding domain regulates p53-mediated apoptosis. J Biol Chem 286:4264–4270. doi:10.1074/jbc.M110.184663

    Article  PubMed  CAS  Google Scholar 

  • Mercurio C, Minucci S, Pelicci PG (2010) Histone deacetylases and epigenetic therapies of hematological malignancies. Pharmacol Res 62:18–34. doi:10.1016/j.phrs.2010.02.010

    Article  PubMed  CAS  Google Scholar 

  • Mérino D, Giam M, Hughes PD, Siggs OM, Heger K, O’Reilly LA, Adams JM, Strasser A, Lee EF, Fairlie WD, Bouillet P (2009) The role of BH3-only protein Bim extends beyond inhibiting Bcl-2-like prosurvival proteins. J Cell Biol 186:355–362. doi:10.1083/jcb.200905153

    Article  PubMed  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and citotoxicity assays. J Immunol Methods 65:55–63. doi:10.1016/0022-1759(83)90303-4

    Article  PubMed  CAS  Google Scholar 

  • Nijhawan D, Fang M, Traer E, Zhong Q, Gao W, Du F, Wang X (2003) Elimination of Mcl-1 is required for the initiation of apoptosis following ultraviolet irradiation. Genes Dev 17:1475–1486. doi:10.1101/gad.1093903

    Article  PubMed  CAS  Google Scholar 

  • O’Connor OA, Heaney ML, Schwartz L, Richardson S, Willim R, MacGregor-Cortelli B, Curly T, Moskowitz C, Portlock C, Horwitz S, Zelenetz AD, Frankel S, Richon V, Marks P, Kelly WK (2006) Clinical experience with intravenous and oral formulations of the novel histone deacetylase inhibitor suberoylanilide hydroxamic acid in patients with advanced hematologic malignancies. J Clin Oncol 24:166–173. doi:10.1200/JCO.2005.01.9679

    Article  PubMed  Google Scholar 

  • Oriol A, Ribera JM, Bergua J, Giménez MesaE, Grande C, Esteve J, Brunet S, Moreno MJ, Escoda L, Hernandez-Rivas JM, Hoelzer D (2008) High-dose chemotherapy and immunotherapy in adult Burkitt lymphoma: comparison of results in human immunodeficiency virus-infected and noninfected patients. Cancer 113:117–125. doi:10.1002/cncr.23522

    Article  PubMed  CAS  Google Scholar 

  • Patte C, Auperin A, Gerrard M, Michon J, Pinkerton R, Sposto R, Weston C, Raphael M, Perkins SL, McCarthy K, Cairo MS (2007) FAB/LMB96 International Study Committee. Results of the randomized international FAB/LMB96 trial for intermediate risk B-cell non-Hodgkin lymphoma in children and adolescents: it is possible to reduce treatment for the early responding patients. Blood 109:2773–2780. doi:10.1182/blood-2006-07-036673

    Google Scholar 

  • Richter-Larrea JA, Robles EF, Fresquet V, Beltran E, Rullan AJ, Agirre X, Calasanz MJ, Panizo C, Richter JA, Hernandez JM, Roman-Gomez J, Prosper F, Martinez-Climent JA (2010) Reversion of epigenetically mediated Bim silencing overcomes chemoresistance in Burkitt lymphoma. Blood 116:2531–2542. doi:10.1182/blood-2010-02-268003

    Article  PubMed  CAS  Google Scholar 

  • Rosato R, Maggio SC, Almenara JA, Payne SG, Atadja P, Spiegel S, Dent P, Grant S (2006) The histone deacetylase inhibitor LAQ824 induces human leukemia cell death through a process involving XIAP down-regulation, oxidative injury, and the acid sphingomyelinase-dependent generation of ceramide. Mol Pharmacol 69:216–225. doi:10.1124/mol.105.017145

    PubMed  CAS  Google Scholar 

  • Sakajiri S, Kumagai T, Kawamata N, Saitoh T, Said JW, Koeffler HP (2005) Histone deacetylase inhibitors profoundly decrease proliferation of human lymphoid cancer cell lines. Exp Hematol 33:53–61. doi:10.1016/j.exphem.2004.09.008

    Article  PubMed  CAS  Google Scholar 

  • Siegel D, Hussein M, Belani C, Robert F, Galanis E, Richon VM, Garcia-Vargas J, Sanz-Rodriguez C, Rizvi S (2009) Vorinostat in solid and hematologic malignancies. J Hematol Oncol 2:31–41. doi:10.1186/1756-8722-2-31

    Article  PubMed  Google Scholar 

  • Silva KL, Vasconcellos DV, Castro EDP, Coelho AM, Linden R, Maia RC (2006) Apoptotic effect of fludarabine is independent of expression of IAPs in B-cell chronic lymphocytic leukaemia. Apoptosis 11:277–285. doi:10.1007/s10495-006-3560-5

    Article  PubMed  CAS  Google Scholar 

  • Tan J, Cang S, Ma Y, Petrillo RL, Liu D (2010) Novel histone deacetylase inhibitors in clinical trials as anti-cancer agents. J Hematol Oncol 4:3–5. doi:10.1186/1756-8722-3-5

    Google Scholar 

  • Thorley-Lawson DA, Allday MJ (2008) The curious case of the tumour virus: 50 years of Burkitt’s lymphoma. Nat Rev Microbiol 6:913–924. doi:10.1038/nrmicro2015

    Article  PubMed  CAS  Google Scholar 

  • Vayssade M, Haddada H, Faridoni-Laurens L, Tourpin S, Valent A, Bénard J, Ahomadegbe JC (2005) P73 functionally replaces p53 in Adriamycin-treated, p53-deficient breast cancer cells. Int J Cancer 116:860–869. doi:10.1002/ijc.21033

    Article  PubMed  CAS  Google Scholar 

  • Walkinshaw DR, Yang XJ (2008) Histone deacetylase inhibitors as novel anticancer therapeutics. Curr Oncol 15:237–243

    PubMed  CAS  Google Scholar 

  • Wang S, Yan-Neale Y, Cai R, Alimov I, Cohen D (2006) Activation of mitochondrial pathway is crucial for tumor selective induction of apoptosis by LAQ824. Cell Cycle 5:1662–1668. doi:10.4161/cc.5.15.3099

    Article  PubMed  CAS  Google Scholar 

  • Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB, Korsmeyer SJ (2001) Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292:727–730. doi:10.1126/science.1059108

    Article  PubMed  CAS  Google Scholar 

  • Xu WS, Parmigiani R, Marks PA (2007) Histone deacetylase inhibitors: molecular mechanisms of action. Oncogene 26:5541–5552. doi:10.1038/sj.onc.1210620

    Article  PubMed  CAS  Google Scholar 

  • Youle J, Strasser A (2008) The Bcl-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9:47–59. doi:10.1038/nrm2308

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Flavia da Cunha Vasconcelos for helping on flow cytometry analysis. This work was supported by a grant from INCT para Controle do Câncer, CNPq 573806/2008-0 e FAPERJ E26/170.026/2008.

Conflict of interest

None of the authors have any financial interest related to this work.

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Authors and Affiliations

  1. Programa de Pesquisa em Hemato-Oncologia Molecular, Coordenação Geral Técnico-Científica, Instituto Nacional de Câncer-INCA, Rio de Janeiro, Brazil

    Ana Carolina dos Santos Ferreira, Renan Amphilophio Fernandes & Jolie Kiemlian Kwee

  2. Programa de Pesquisa em Hemato-Oncologia Molecular, Coordenação Geral Técnico-Científica, Instituto Nacional de Câncer, Praça Cruz Vermelha, 23/6th floor, Rio de Janeiro, CEP 20 230-130, Brazil

    Claudete Esteves Klumb

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  1. Ana Carolina dos Santos Ferreira
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Correspondence to Claudete Esteves Klumb.

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dos Santos Ferreira, A.C., Fernandes, R.A., Kwee, J.K. et al. Histone deacetylase inhibitor potentiates chemotherapy-induced apoptosis through Bim upregulation in Burkitt’s lymphoma cells. J Cancer Res Clin Oncol 138, 317–325 (2012). https://doi.org/10.1007/s00432-011-1093-y

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