Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Pharmacological inhibitors of NF-κB accelerate apoptosis in chronic lymphocytic leukaemia cells

Oncogene volume 26, pages 1166–1177 (2007)Cite this article

Abstract

Nuclear factor-kappaB (NF-κB) is a transcription factor that plays a critical role in the inappropriate survival of various types of malignant cells. Chronic lymphocytic leukaemia (CLL) is the most common B-cell malignancy in the Western world. Although overexpression and regulation of NF-κB has been described in CLL, its function remains unclear. Exposure of CLL cells to BAY117082 or Kamebakaurin, potent pharmacological inhibitors of the NF-κB pathway, accelerated apoptosis in approximately 70% of cases. Sensitivity to NF-κB pathway inhibitors was not related to the prognostic markers VH status, CD38 or Zap70 expression, or to the levels of nuclear NF-κB. Normal peripheral B cells were resistant to the apoptosis-inducing effects of these compounds. Cell death induced by the inhibitors was associated with activation of caspase-9 and -3, and loss of mitochondrial membrane polarization, but did not involve changes in the expression of Bcl-2 or Mcl-1. Inhibitors caused an increase in c-jun NH2-terminal kinase activity in CLL, but this did not appear to be important for apoptosis. Microarray analysis identified some potential novel NF-κB target genes, including interleukin-16- and the Bcl-2- related survival protein Bcl-w. These results demonstrate that a substantial proportion of CLL are dependent on NF-κB for enhanced survival and suggest that inhibition of NF-κB may have therapeutic potential.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  • Bernal A, Pastore RD, Asgary Z, Keller SA, Cesarman E, Liou HC et al. (2001). Survival of leukemic B cells promoted by engagement of the antigen receptor. Blood 98: 3050–3057.

    Article  CAS  PubMed  Google Scholar 

  • Brenner C, Le Bras M, Kroemer G . (2003). Insights into the mitochondrial signaling pathway: what lessons for chemotherapy? J Clin Immunol 23: 73–80.

    Article  CAS  PubMed  Google Scholar 

  • Caligaris-Cappio F, Hamblin TJ . (1999). B-cell chronic lymphocytic leukemia: a bird of a different feather. J Clin Oncol 17: 399–408.

    Article  CAS  PubMed  Google Scholar 

  • Castro JE, Prada CE, Loria O, Kamal A, Chen L, Burrows FJ et al. (2005). ZAP-70 is a novel conditional heat shock protein 90 (Hsp90) client inhibition of Hsp90 leads to ZAP-70 degradation, apoptosis, and impaired signaling in chronic lymphocytic leukemia. Blood 106: 2506–2512.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen L, Apgar J, Huynh L, Dicker F, Giago-McGahan T, Rassenti L et al. (2005). ZAP-70 directly enhances IgM signaling in chronic lymphocytic leukemia. Blood 105: 2036–2041.

    Article  CAS  PubMed  Google Scholar 

  • Claudio E, Brown K, Park S, Wang H, Siebenlist U . (2002). BAFF-induced NEMO-independent processing of NF-kappa B2 in maturing B cells. Nat Immunol 3: 958–965.

    Article  CAS  PubMed  Google Scholar 

  • Crespo M, Bosch F, Villamor N, Bellosillo B, Colomer D, Rozman M et al. (2003). ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. N Engl J Med 348: 764–775.

    Article  Google Scholar 

  • Cuni S, Perez-Aciego P, Perez-Chacon G, Vargas JA, Sanchez A, Martin-Saavedra FM et al. (2004). A sustained activation of P13K/NF-kappaB pathway is critical for the survival of chronic lymphocytic leukemia B cells. Leukemia 18: 1391–1400.

    Article  CAS  PubMed  Google Scholar 

  • Dallman C, Packham G . (2005). Purification of primary malignant B-cells and immunoblot analysis of bcl-2 family proteins. Methods Mol Med 115: 1–13.

    CAS  PubMed  Google Scholar 

  • Damle RN, Wasil T, Fais F, Ghiotto F, Valetto A, Allen SL et al. (1999). IgVgene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood 94: 1840–1847.

    CAS  PubMed  Google Scholar 

  • DeMeester SL, Buchman TG, Qiu Y, Dunnigan K, Hotchkiss RS, Karl IE et al. (1998). Pyrrolidine dithiocarbamate activates the heat shock response and thereby induces apoptosis in primed endothelial cells. Shock 10: 1–6.

    Article  CAS  PubMed  Google Scholar 

  • Feuillard J, Schuhmacher M, Kohanna S, Asso-Bonnet M, Ledeur F, Joubert-Caron R et al. (2000). Inducible loss of NF-kappaB activity is associated with apoptosis and Bcl-2 down-regulation in Epstein–Barr virus-transformed B lymphocytes. Blood 95: 2068–2075.

    CAS  PubMed  Google Scholar 

  • Furman RR, Asgary Z, Mascarenhas JO, Liou HC, Schattner EJ . (2000). Modulation of NF-kappa B activity and apoptosis in chronic lymphocytic leukemia B cells. J Immunol 164: 2200–2206.

    Article  CAS  PubMed  Google Scholar 

  • Ghia P, Caligaris-Cappio F . (2000). The indispensable roll of microenvironment in the natural history of low-grade B-cell neoplasms. Adv Cancer Res 79: 157–173.

    Article  CAS  PubMed  Google Scholar 

  • Grad JM, Zeng XR, Boise LH . (2000). Regulation of Bcl-xL: a little bit of this and a little bit of STAT. Curr Opin Oncol 12: 543–549.

    Article  CAS  PubMed  Google Scholar 

  • Granziero L, Ghia P, Circosta P, Gottardi D, Strola G, Geuna M et al. (2001). Survivin is expressed on CD40 stimulation and interfaces proliferation and apoptosis in B-cell chronic lymphocytic leukemia. Blood 97: 2777–2783.

    Article  CAS  PubMed  Google Scholar 

  • Hamblin TJ, Davis Z, Gardiner A, Oscier DG, Stevenson FK . (1999). Unmutated Ig V(H) genes are associated with a more aggressive form of chronic lymphocytic leukemia. Blood 94: 1848–1854.

    CAS  PubMed  Google Scholar 

  • Hamblin TJ, Orchard JA, Ibbotson RE, Davis Z, Thomas PW, Stevenson FK et al. (2002). CD38 expression and immunoglobulin variable region mutations are independent prognostic variables in chronic lymphocytic leukemia, but CD38 expression may vary during the course of the disease. Blood 99: 1023–1029.

    Article  CAS  PubMed  Google Scholar 

  • Hess P, Pihan G, Sawyers CL, Flavell RA, Davis RJ . (2002). Survival signaling mediated by c-Jun NH(2)-terminal kinase in transformed B lymphoblasts. Nat Genet 32: 201–205.

    Article  CAS  PubMed  Google Scholar 

  • Kayagaki N, Yan M, Seshasayee D, Wang H, Lee W, French DM et al. (2002). BAFF/BlyS receptor 3 binds the B cell survival factor BAFF ligand through a discrete surface loop and promotes processing of NF-KappaB2. Immunity 17: 515–524.

    Article  CAS  PubMed  Google Scholar 

  • Kern C, Cornuel JF, Billard C, Tang R, Rouillard D, Stenou V et al. (2004). Involvement of BAFF and APRIL in the resistance to apoptosis of B-CLL through an autocrine pathway. Blood 103: 679–688.

    Article  CAS  PubMed  Google Scholar 

  • LaCasse EC, Baird S, Korneluk RG, MacKenzie AE . (1998). The inhibitors of apoptosis (LAPs) and their emerging role in cancer. Oncogene 17: 3247–3259.

    Article  PubMed  Google Scholar 

  • Lee HH, Dadgostar H, Cheng Q, Shu J, Cheng G . (1999). NF-kappaB-mediated up-regulation of Bcl-x and Bfl-1/A1 is required for CD40 survival signaling in B lymphocytes. Proc Natl Acad Sci USA 96: 9136–9141.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee JH, Koo TH, Hwang BY, Lee JJ . (2002). Kaurane diterpene, kamebakaurin, inhibits NF-kappa B by directly targeting the DNA-binding activity of p50 andblocks the expression of antiapoptotic NF-kappa B target genes. J Biol Chem 277: 18411–18420.

    Article  CAS  PubMed  Google Scholar 

  • Matsuda A, Suzuki Y, Honda G, Muramatsu S, Matsuzaki O, Nagano Y et al. (2003). Large-scale identification and characterization of human genes that activate NF-kappaB and MARK signaling pathways. Oncogene 22: 3307–3318.

    Article  CAS  PubMed  Google Scholar 

  • Michels J, O’Neill JW, Dallman CL, Mouzakiti A, Habens F, Brimmell M et al. (2004). Ncl-1 is required for Akata6 B-lymphoma cell survival and is converted to a cell death molecule by efficient caspase-mediated cleavage. Oncogene 23: 4818–4827.

    Article  CAS  PubMed  Google Scholar 

  • Morabito F, Mangiola M, Oliva B, Stelitano C, Callea V, Deaglio S et al. (2001). Peripheral blood CD38 expression predicts survival in B-cell chronic lymphocytic leukemia. Leukemia Res 25: 927–932.

    Article  CAS  Google Scholar 

  • Munzert G, Kirchner D, Stobbe H, Bergmann L, Schmid RM, Dohner H . (2002). Tumour necrosis factor receptor-associated factor 1 gene overexpression in B-cell chronic lymphocytic leukemia: analysis of NF-kappa B/Rel-regulated inhibitors of apoptosis. Blood 100: 3749–3756.

    Article  CAS  PubMed  Google Scholar 

  • Natoli G, Saccani S, Bosisio D, Marazzi I . (2005). Interactions of NF-kappaB with chromatin: the art of being at the right place at the right time. Nat Immunol 6: 439–445.

    Article  CAS  PubMed  Google Scholar 

  • Oakley F, Mann J, Ruddell RG, Pickford J, Weinmaster G, Mann DA . (2003). Basal expression of IkappaBalpha is controlled by the mammalian transcriptional repressor RBP-J (CBF1) and its activator Notch1. J Biol Chem 278: 24359–24370.

    Article  CAS  PubMed  Google Scholar 

  • Orlowski RZ, Baldwin Jr AS . (2002). NF-kappa B as a therapeutic target in cancer. Trends Mol Med 8: 385–389.

    Article  CAS  PubMed  Google Scholar 

  • Packham G, Stevenson FK . (2005). Bodyguards and assassins: Bcl-2 family proteins and apoptosis control in chronic lymphocytic leukaemia. Immunology 114: 441–449.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Packham G, Brimmell M, Cleveland JL . (1997). Mammalian cells express two differently localized Bag-1 isoforms generated by alternative translation initiation. Biochem J 328: 807–813.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Papa S, Zazzeroni F, Pham CG, Bubici C, Franzoso G . (2004). Linking JNK signaling to NF-kappaB: a key to survival. J Cell Sci 117: 5197–5208.

    Article  CAS  PubMed  Google Scholar 

  • Pedersen IM, Kitada S, Leoni LM, Zapata JM, Karras JG, Tsukada N et al. (2002). Protection of CLL B cells by a follicular dendritic cell line is dependent on induction of Mcl-1. Blood 100: 1795–1801.

    Article  CAS  PubMed  Google Scholar 

  • Pepper C, Thomas A, Hoy T, Cotter F, Bentley P . (1999). Antisense-mediated suppression of Bcl-2 highlights its pivotol role in failed apoptosis in B-cell chronic lymphocytic leukaemia. Br J Haematol 107: 611–615.

    Article  CAS  PubMed  Google Scholar 

  • Petlickovski A, Laurenti L, Li X, Marietti S, Chiusolo P, Sica S et al. (2005). Sustained signaling through the B-cell receptor induces Mcl-1 and promotes survival of chronic lymphocytic leukemia B cells. Blood 105: 4820–4827.

    Article  CAS  PubMed  Google Scholar 

  • Perkins ND . (2004). NF-kappa B: tumour promoter or suppressor? Trends Cell Biol 14: 64–69.

    Article  CAS  PubMed  Google Scholar 

  • Pierce JW, Schoenleber R, Jesmok G, Best J, Moore SA, Collins T et al. (1997). Novel inhibitors of cytokine-induced Ikappa Balpha phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo. J Biol Chem 272: 12103–21096.

    Google Scholar 

  • Ran R, Lu A, Zhang L, Tang Y, Zhu H, Xu H et al. (2004). Hsp70 promotes TNF-mediated apoptosis by binding IKK, gamma and impairing NF-kappaB survival signaling. Genes Dev 18: 1466–1481.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ravid T, Hochstrasser M . (2004). NF-kappaB signaling flipping the switch with polyubiquitin chains. Curr Biol 14: R898–R900.

    Article  CAS  PubMed  Google Scholar 

  • Ringshausen I, Schneller F, Bogner C, Hipp S, Duyster J, Peschel C et al. (2002). Constitutively activated phosphatidylinositol-3 kinase (PI-3K) is involved in the defect of apoptosis in B-CLL association with protein kinase Cdelta. Blood 100: 3741–3748.

    Article  CAS  PubMed  Google Scholar 

  • Rodriguez A, Martinez N, Camacho FI, Ruiz-Ballesteros E, Algara P, Garcia JF et al. (2004). Variability in the degree of expression of phosphorylated Ikappa Balpha in chronic lymphocytic leukemia cases with nodial involvement. Clin Cancer Res 10: 6796–6806.

    Article  CAS  PubMed  Google Scholar 

  • Romano MF, Lamberti A, Tassone P, Alfinito F, Costantini S, Chiurazzi F et al. (1998). Triggering of CD40 antigen inhibits fludarabine-induced apoptosis in B chronic lymphocytic leukemia cells. Blood 92: 990–995.

    CAS  PubMed  Google Scholar 

  • Rosenwald A, Alizadeh AA, Widhopf G, Simon R, Davis RE, Yu X et al. (2001). Relation of gene expression phenotype to immunoglobulin mutation genotype in Bcell chronic lymphocytic leukemia. J Exp Med 194: 1639–1647.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rossi A, Elia G, Santoro MG . (1998). Activation of the heatshock factor 1 by serine protease inhibitors. An effect associated with nuclear factor-kappaB inhibition. J Biol Chem 273: 16446–16452.

    Article  CAS  PubMed  Google Scholar 

  • Sembries S, Pahl H, Stilgenbauer S, Dohner H, Schriever F . (1999). Reduced expression of adhesion molecules andcell signaling receptors by chronic lymphocytic leukemia cells with 11q deletion. Blood 93: 624–631.

    CAS  PubMed  Google Scholar 

  • Shaffer AL, Rosenwald A, Staudt LM . (2002). Lymphoid malignancies: the dark side of B-cell differentiation. Nat Rev Immunol 2: 920–932.

    Article  CAS  PubMed  Google Scholar 

  • Schattner EJ . (2000). CD40 ligand in CLL pathogenesis and therapy. Leukemia Lymphoma 37: 461–472.

    Article  CAS  PubMed  Google Scholar 

  • Schmitz ML, Mattioli I, Buss H, Kracht M . (2004). NF-kappaB: a multifaceted transcription factor regulated at several levels. Chembiochem 5: 348–358.

    Article  Google Scholar 

  • Scorrano L, Korsmeyer SJ . (2003). Mechanisms of cytochrome c release by proapoptotic BCL-2 family members. Biochem Biophys Res Commun 304: 437–444.

    Article  CAS  PubMed  Google Scholar 

  • Takayama S, Reed JC . (2001). Molecular chaperone targeting and regulation by BAG family proteins. Nat Cell Biol 3: E237–E241.

    Article  CAS  PubMed  Google Scholar 

  • Tran NL, McDonough WS, Savitch BA, Sawyer TF, Winkles JA, Berens ME . (2005). The tumour necrosis factor-like weak inducer of apoptosis (TWEAK)-fibroblast growth factor-inducible 14(Fn14) signaling system regualtes glioma cell survival via NfkappaB pathway activiation and BCL-XL-W expression. J Biol Chem 280: 3483–3492.

    Article  CAS  PubMed  Google Scholar 

  • Viatour P, Bentires-Alj M, Chariot A, Deregowski V, de Leval L, Merville MP et al. (2003). NF-kappa B2/p100 induces Bcl-2 expression. Leukemia 17: 1349–1356.

    Article  CAS  PubMed  Google Scholar 

  • Wiestner A, Rosenwald A, Barry TS, Wright G, Davis RE, Henrickson SE et al. (2003). ZAP-70 expression identifies a chronic lymphocytic leukemia sub-type with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood 101: 4944–4951.

    Article  CAS  PubMed  Google Scholar 

  • Wilson KC, Center DM, Cruikshank WW . (2004). The effect of interleukin-16 and its precursor on T-lymphocyte activation and growth. Growth Factors 22: 97–104.

    Article  CAS  PubMed  Google Scholar 

  • Zamzami N, Marchetti P, Castedo M, Zanin C, Vayssiere JL, Petit PX et al. (1995). Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo. J Exp Med 181: 1661–1672.

    Article  CAS  PubMed  Google Scholar 

  • Zaninoni A, Imperiali FG, Pasquini C, Zanella A, Barcellini W . (2003). Cytokkine modulation of nuclear factor-kappaB activity in B-chronic lymphocytic leukemia. Exp Hematol 31: 185–190.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Isla Wheatley and Dr C Ian Mockridge for technical assistance and the following clinical collaborators for kindly providing material: Dr Andrew Duncombe and Dr Debbie Richardson of the Southampton General Hospital, Southampton; Professor Terry Hamblin of the Royal Bournemouth Hospital, Bournemouth, UK, Dr Abraham Jacob of the New Cross Hospital, Wolverhampton and Dr H McCarthy of the Queen Alexandra Hospital, Portsmouth. This work was funded by Leukaemia Research Fund, Tenovus and Cancer Research UK.

Author information

Authors and Affiliations

  1. Cancer Research UK Clinical Centre, Southampton General Hospital, Southampton, UK

    B M Pickering, S de Mel, M Lee, M Howell, F Habens, C L Dallman, P W M Johnson & G Packham

  2. Cancer Sciences Division, Molecular Immunology Group, Southampton General Hospital, Southampton, UK

    L A Neville, K N Potter & F K Stevenson

  3. Division of Infection, Liver Group, Inflammation and Repair, University of Southampton School of Medicine, Southampton General Hospital, Southampton, UK

    J Mann & D A Mann

Authors
  1. B M Pickering
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S de Mel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M Howell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. F Habens
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C L Dallman
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. L A Neville
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. K N Potter
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. J Mann
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. D A Mann
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. P W M Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. F K Stevenson
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. G Packham
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G Packham.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pickering, B., de Mel, S., Lee, M. et al. Pharmacological inhibitors of NF-κB accelerate apoptosis in chronic lymphocytic leukaemia cells. Oncogene 26, 1166–1177 (2007). https://doi.org/10.1038/sj.onc.1209897

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1209897

Keywords

This article is cited by

Search

Advanced search

Quick links