Expression of multidrug resistance-associated ABC transporters in B-CLL is independent of ZAP70 status

  • Alexander Hoellein
  • Thomas Decker
  • Christian Bogner
  • Madlen Oelsner
  • Stefanie Hauswald
  • Christian Peschel
  • Ulrich Keller
  • Thomas Licht
Original Paper



To assess whether the poor prognosis of ZAP70-positive B-cell chronic lymphocytic leukemia (CLL) is associated with the overexpression of ABC transporter genes that are responsible for pleiotropic drug resistance.

Materials and methods

The transcript level of ten drug transporters was analyzed using semiquantitative and quantitative RT-PCR in control hematopoietic cells, in 41 CLL patient samples and in 5 lymphoma cell lines. ZAP70 status was determined by immunoblotting.


Of all analyzed transporters, MDR1, MDR2, MRP1, MRP4, MRP5, and MRP7 were expressed at a significantly higher level in B lymphocytes when compared with other hematopoietic cells in peripheral blood. A subgroup of 41 CLL patient samples showed similar or higher expression of these genes than control B cells, and CLL cells exhibited high expression when compared with multiple lymphoma cell lines. No significant correlation between ZAP70 expression and ABC transporter expression was observed.


The ZAP70 status is independent of the multidrug resistance phenotype in CLL.


CLL ABC transporter Multidrug resistance ZAP70 



This work was supported by the German José Carreras Leukemia Foundation (Project R04/17 to T. Licht and U. Keller). We are grateful to M. Gottesman, National Cancer Institute, Bethesda, USA, for providing the drug-resistant cell lines KB-3-1 and KB-8-5. We also would like to thank T. Schuster, Institute for Medical Statistics and Epidemiology, Technische Universität München, Munich, Germany, for help with statistical analysis.

Conflict of interest statement

We declare that we have no conflict of interest.


  1. Andreadis C, Gimotty PA, Wahl P, Hammond R, Houldsworth J, Schuster SJ, Rebbeck TR (2007) Members of the glutathione and ABC-transporter families are associated with clinical outcome in patients with diffuse large B-cell lymphoma. Blood 109:3409–3416CrossRefPubMedGoogle Scholar
  2. Bogner C, Sandherr M, Perker M, Weick K, Ringshausen I, Peschel C, Decker T (2006) Cyclin E but not bcl-2, bax or mcl-1 is differentially expressed in ZAP 70-positive and ZAP 70-negative B-CLL cells. Ann Hematol 85:458–462CrossRefPubMedGoogle Scholar
  3. Bosanquet AG, Johnson SA, Richards SM (1999) Prognosis for fludarabine therapy of chronic lymphocytic leukaemia based on ex vivo drug response by DiSC assay. Br J Haematol 106:71–77CrossRefPubMedGoogle Scholar
  4. Chaudhary PM, Roninson IB (1991) Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells. Cell 66:85–94CrossRefPubMedGoogle Scholar
  5. Chen L, Widhopf G, Huynh L, Rassenti L, Rai KR, Weiss A, Kipps TJ (2002) Expression of ZAP-70 is associated with increased B-cell receptor signaling in chronic lymphocytic leukemia. Blood 100:4609–4614CrossRefPubMedGoogle Scholar
  6. Crespo M, Bosch F, Villamor N, Bellosillo B, Colomer D, Rozman M, Marce S, Lopez-Guillermo A, Campo E, Montserrat E (2003) ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. N Engl J Med 348:1764–1775CrossRefPubMedGoogle Scholar
  7. Damle RN, Ghiotto F, Valetto A, Albesiano E, Fais F, Yan XJ, Sison CP, Allen SL, Kolitz J, Schulman P, Vinciguerra VP, Budde P, Frey J, Rai KR, Ferrarini M, Chiorazzi N (2002) B-cell chronic lymphocytic leukemia cells express a surface membrane phenotype of activated, antigen-experienced B lymphocytes. Blood 99:4087–4093CrossRefPubMedGoogle Scholar
  8. Decker T, Oelsner M, Kreitman RJ, Salvatore G, Wang QC, Pastan I, Peschel C, Licht T (2004) Induction of caspase-dependent programmed cell death in B-cell chronic lymphocytic leukemia by anti-CD22 immunotoxins. Blood 103:2718–2726CrossRefPubMedGoogle Scholar
  9. Del Principe MI, Del Poeta G, Buccisano F, Maurillo L, Venditti A, Zucchetto A, Marini R, Niscola P, Consalvo MA, Mazzone C, Ottaviani L, Panetta P, Bruno A, Bomben R, Suppo G, Degan M, Gattei V, de Fabritiis P, Cantonetti M, Lo Coco F, Del Principe D, Amadori S (2006) Clinical significance of ZAP-70 protein expression in B-cell chronic lymphocytic leukemia. Blood 108:853–861CrossRefPubMedGoogle Scholar
  10. Fojo AT, Ueda K, Slamon DJ, Poplack DG, Gottesman MM, Pastan I (1987) Expression of a multidrug-resistance gene in human tumors and tissues. Proc Natl Acad Sci USA 84:265–269CrossRefPubMedGoogle Scholar
  11. Goldstein LJ, Galski H, Fojo A, Willingham M, Lai SL, Gazdar A, Pirker R, Green A, Crist W, Brodeur GM (1989) Expression of a multidrug resistance gene in human cancers. J Natl Cancer Inst 81:116–124CrossRefPubMedGoogle Scholar
  12. Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58CrossRefPubMedGoogle Scholar
  13. Guillaume N, Gouilleux-Gruart V, Claisse JF, Troussard X, Lepelley P, Damaj G, Royer B, Garidi R, Lefrere JJ (2007) Multi-drug resistance mediated by P-glycoprotein overexpression is not correlated with ZAP-70/CD38 expression in B-cell chronic lymphocytic leukemia. Leuk Lymphoma 48:1556–1560CrossRefPubMedGoogle Scholar
  14. Guo Y, Kotova E, Chen ZS, Lee K, Hopper-Borge E, Belinsky MG, Kruh GD (2003) MRP8, ATP-binding cassette C11 (ABCC11), is a cyclic nucleotide efflux pump and a resistance factor for fluoropyrimidines 2′, 3′-dideoxycytidine and 9′-(2′-phosphonylmethoxyethyl)adenine. J Biol Chem 278:29509–29514CrossRefPubMedGoogle Scholar
  15. Hallek M, Cheson BD, Catovsky D, Caligaris-Cappio F, Dighiero G, Dohner H, Hillmen P, Keating MJ, Montserrat E, Rai KR, Kipps TJ (2008) Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia (IWCLL) updating the National Cancer Institute-Working Group (NCI-WG) 1996 guidelines. Blood 111:5446–5456CrossRefPubMedGoogle Scholar
  16. 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 94:1848–1854PubMedGoogle Scholar
  17. Herweijer H, Sonneveld P, Baas F, Nooter K (1990) Expression of mdr1 and mdr3 multidrug-resistance genes in human acute and chronic leukemias and association with stimulation of drug accumulation by cyclosporine. J Natl Cancer Inst 82:1133–1140CrossRefPubMedGoogle Scholar
  18. Jerkeman M, Anderson H, Dictor M, Kvaloy S, Akerman M, Cavallin-Stahl E (2004) Assessment of biological prognostic factors provides clinically relevant information in patients with diffuse large B-cell lymphoma—a Nordic Lymphoma Group study. Ann Hematol 83:414–419CrossRefPubMedGoogle Scholar
  19. Matthews C, Catherwood MA, Larkin AM, Clynes M, Morris TC, Alexander HD (2006) MDR-1, but not MDR-3 gene expression, is associated with unmutated IgVH genes and poor prognosis chromosomal aberrations in chronic lymphocytic leukemia. Leuk Lymphoma 47:2308–2313CrossRefPubMedGoogle Scholar
  20. Molina-Arcas M, Bellosillo B, Casado FJ, Montserrat E, Gil J, Colomer D, Pastor-Anglada M (2003) Fludarabine uptake mechanisms in B-cell chronic lymphocytic leukemia. Blood 101:2328–2334CrossRefPubMedGoogle Scholar
  21. Nooter K, Sonneveld P, Janssen A, Oostrum R, Boersma T, Herweijer H, Valerio D, Hagemeijer A, Baas F (1990) Expression of the mdr3 gene in prolymphocytic leukemia: association with cyclosporin-A-induced increase in drug accumulation. Int J Cancer 45:626–631CrossRefPubMedGoogle Scholar
  22. Ohsawa M, Ikura Y, Fukushima H, Shirai N, Sugama Y, Suekane T, Hirayama M, Hino M, Ueda M (2005) Immunohistochemical expression of multidrug resistance proteins as a predictor of poor response to chemotherapy and prognosis in patients with nodal diffuse large B-cell lymphoma. Oncology 68:422–431CrossRefPubMedGoogle Scholar
  23. Pepper C, Thomas A, Hoy T, Bentley P (1999) Chlorambucil resistance in B-cell chronic lymphocytic leukaemia is mediated through failed Bax induction and selection of high Bcl-2-expressing subclones. Br J Haematol 104:581–588CrossRefPubMedGoogle Scholar
  24. Ritzel MW, Ng AM, Yao SY, Graham K, Loewen SK, Smith KM, Ritzel RG, Mowles DA, Carpenter P, Chen XZ, Karpinski E, Hyde RJ, Baldwin SA, Cass CE, Young JD (2001) Molecular identification and characterization of novel human and mouse concentrative Na+-nucleoside cotransporter proteins (hCNT3 and mCNT3) broadly selective for purine and pyrimidine nucleosides (system cib). J Biol Chem 276:2914–2927CrossRefPubMedGoogle Scholar
  25. Rosenwald A, Alizadeh AA, Widhopf G, Simon R, Davis RE, Yu X, Yang L, Pickeral OK, Rassenti LZ, Powell J, Botstein D, Byrd JC, Grever MR, Cheson BD, Chiorazzi N, Wilson WH, Kipps TJ, Brown PO, Staudt LM (2001) Relation of gene expression phenotype to immunoglobulin mutation genotype in B cell chronic lymphocytic leukemia. J Exp Med 194:1639–1647CrossRefPubMedGoogle Scholar
  26. Schaich M, Soucek S, Thiede C, Ehninger G, Illmer T (2005) MDR1 and MRP1 gene expression are independent predictors for treatment outcome in adult acute myeloid leukaemia. Br J Haematol 128:324–332CrossRefPubMedGoogle Scholar
  27. Shen DW, Cardarelli C, Hwang J, Cornwell M, Richert N, Ishii S, Pastan I, Gottesman MM (1986) Multiple drug-resistant human KB carcinoma cells independently selected for high-level resistance to colchicine, adriamycin, or vinblastine show changes in expression of specific proteins. J Biol Chem 261:7762–7770PubMedGoogle Scholar
  28. Sonneveld P, Nooter K, Burghouts JT, Herweijer H, Adriaansen HJ, van Dongen JJ (1992) High expression of the mdr3 multidrug-resistance gene in advanced-stage chronic lymphocytic leukemia. Blood 79:1496–1500PubMedGoogle Scholar
  29. Tangye SG, Raison RL (1997) Human cytokines suppress apoptosis of leukaemic CD5+ B cells and preserve expression of bcl-2. Immunol Cell Biol 75:127–135CrossRefPubMedGoogle Scholar
  30. Wiestner A, Rosenwald A, Barry TS, Wright G, Davis RE, Henrickson SE, Zhao H, Ibbotson RE, Orchard JA, Davis Z, Stetler-Stevenson M, Raffeld M, Arthur DC, Marti GE, Wilson WH, Hamblin TJ, Oscier DG, Staudt LM (2003) ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood 101:4944–4951CrossRefPubMedGoogle Scholar
  31. Zent CS, Call TG, Hogan WJ, Shanafelt TD, Kay NE (2006) Update on risk-stratified management for chronic lymphocytic leukemia. Leuk Lymphoma 47:1738–1746CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Alexander Hoellein
    • 1
  • Thomas Decker
    • 1
  • Christian Bogner
    • 1
  • Madlen Oelsner
    • 1
  • Stefanie Hauswald
    • 1
  • Christian Peschel
    • 1
  • Ulrich Keller
    • 1
  • Thomas Licht
    • 1
    • 2
  1. 1.III Medical DepartmentTechnische Universität MünchenMunichGermany
  2. 2.HELIOS SchlossbergklinikOberstaufenGermany

Personalised recommendations