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Epigenetic Regulation of CD20 Protein Expression in a Novel B-Cell Lymphoma Cell Line, RRBL1, Established from a Patient Treated Repeatedly with Rituximab-Containing Chemotherapy

International Journal of Hematology volume 86, pages 49–57 (2007)Cite this article

Abstract

Rituximab is a chimeric monoclonal antibody to the surface antigen CD20 and has provided better outcomes against CD20+ B-cell lymphomas than chemotherapy with conventional antitumor drugs alone. Treatment with rituximab poses a considerable problem, however, because of CD20- tumor transformation and subsequent disease progression. We have established a CD20- lymphoma cell line, RRBL1, from a diffuse large B-cell lymphoma with CD20- transformation from CD20+ follicular lymphoma after treatment with rituximab. RRBL1 was CD10+, CD19+, and CD20- by flow cytometry. CD20 expression was not detected by immunohistochemistry. Immunoblotting with whole RRBL1 cell lysate showed a very faint CD20 band only with longer exposures. The level of CD20 messenger RNA (mRNA) expression detected by quantitative reverse transcriptase-polymerase chain reaction analysis was almost 100 times lower than that in CD20+ lymphoma cells. When we treated RRBL1 cells with trichostatin A, an epigenetic drug that modulates histone-acetylation status, we detected dramatically increased CD20 mRNA and protein expression, suggesting that epigenetic mechanisms may explain the CD20- phenotype in RRBL1 cells. Thus, RRBL1 may be useful not only for analyses of mechanisms for the absence of CD20 expression in vitro but also for exploration of therapies against CD20- B-cell malignancies in vivo.

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References

  1. Cheson BD. Monoclonal antibody therapy for B-cell malignancies.Semin Oncol. 2006;33:S2-S14.

    Article  CAS  Google Scholar 

  2. Imai K, Takaoka A. Comparing antibody and small-molecule therapies for cancer.Nat Rev Cancer. 2006;6:714–727.

    Article  CAS  Google Scholar 

  3. Coiffier B, Lepage E, Briere J, et al. CHOP chemotherapy plus rituximab compared with CHOP alone in elderly patients with diffuse large-B-cell lymphoma.N Engl J Med. 2002;346:235–242.

    Article  PubMed  CAS  Google Scholar 

  4. van OersMH, Klasa R, Marcus RE, et al. Rituximab maintenance improves clinical outcome of relapsed/resistant follicular non-Hodgkin’s lymphoma both in patients with and without rituximab during induction: results of a prospective randomized phase III intergroup trial.Blood. 2006;108:3295–3301.

    Article  PubMed  CAS  Google Scholar 

  5. HabermannTM, Weller EA, Morrison VA, et al. Rituximab-CHOP versus CHOP alone or with maintenance rituximab in older patients with diffuse large B-cell lymphoma.J Clin Oncol. 2006;24:3121–3127.

    Article  CAS  Google Scholar 

  6. Pfreundschuh M, Trumper L, Osterborg A, et al. CHOP-like chemotherapy plus rituximab versus CHOP-like chemotherapy alone in young patients with good-prognosis diffuse large-B-cell lymphoma: a randomised controlled trial by the MabThera International Trial (MInT) Group.Lancet Oncol. 2006;7:379–391.

    Article  PubMed  CAS  Google Scholar 

  7. Hiddemann W, Kneba M, Dreyling M, et al. Frontline therapy with rituximab added to the combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) significantly improves the outcome for patients with advanced-stage follicular lymphoma compared with therapy with CHOP alone: results of a prospective randomized study of the German Low-Grade Lymphoma Study Group.Blood. 2005;106:3725–3732.

    Article  PubMed  CAS  Google Scholar 

  8. Czuczman MS,Weaver R, Alkuzweny B, Berlfein J, Grillo-Lopez AJ. Prolonged clinical and molecular remission in patients with low-grade or follicular non-Hodgkin’s lymphoma treated with rituximab plus CHOP chemotherapy: 9-year follow-up.J Clin Oncol. 2004;22:4711–4716.

    Article  PubMed  CAS  Google Scholar 

  9. Alvaro-Naranjo T, Jaen-Martinez J, Guma-Padro J, Bosch-Princep R, Salvado-Usach MT. CD20-negative DLBCL transformation after rituximab treatment in follicular lymphoma: a new case report and review of the literature.Ann Hematol. 2003; 82:585–588.

    Article  PubMed  CAS  Google Scholar 

  10. Terui Y, Sakurai T, Mishima Y, et al. Identification of CD20 mutations in malignant lymphoma: can they be predictors of response to rituximab? [abstract].Blood. 2005;106:88a. Abstract 288.

    Google Scholar 

  11. Davis TA, Czerwinski DK, Levy R. Therapy of B-cell lymphoma with anti-CD20 antibodies can result in the loss of CD20 antigen expression.Clin Cancer Res. 1999;5:611–615.

    CAS  Google Scholar 

  12. Kinoshita T, Nagai H, Murate T, Saito H. CD20-negative relapse in B-cell lymphoma after treatment with rituximab.J Clin Oncol. 1998;16:3916.

    Article  CAS  Google Scholar 

  13. Smith MR. Rituximab (monoclonal anti-CD20 antibody): mechanisms of action and resistance.Oncogene. 2003;22:7359–7368.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  14. Ninomiya M, Abe A, Yokozawa T, et al. Establishment of a myeloid leukemia cell line, TRL-01, withMLL-ENL fusion gene.Cancer Genet Cytogenet. 2006;169:1–11.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  15. Mason DY, Comans-Bitter WM, Cordell JL, Verhoeven MA, van DongenJJ. Antibody L26 recognizes an intracellular epitope on the B-cell-associated CD20 antigen.Am J Pathol. 1990;136:1215–1222.

    PubMed  PubMed Central  CAS  Google Scholar 

  16. Atsumi A, Tomita A, Kiyoi H, Naoe T. Histone deacetylase 3 (HDAC3) is recruited to target promoters by PML-RARα as a component of the N-CoR co-repressor complex to repress transcriptionin vivo.Biochem Biophys Res Commun. 2006;345:1471–1480.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Kiyoi H, Naoe T, Kitamura K, Yamauchi T, Ichihashi T, Ohno R. Disappearance of minimal residual lymphoblastic leukemia cells 6 months after allogeneic bone marrow transplantation without GVHD.Bone Marrow Transplant. 1991;8:59–62.

    PubMed  PubMed Central  CAS  Google Scholar 

  18. Gribben JG, Freedman A, Woo SD, et al. All advanced stage non-Hodgkin’s lymphomas with a polymerase chain reaction amplifiable breakpoint of bcl-2 have residual cells containing the bcl-2 rearrangement at evaluation and after treatment.Blood. 1991;78:3275–3280.

    PubMed  PubMed Central  CAS  Google Scholar 

  19. Ravetch JV, Siebenlist U, Korsmeyer S, Waldmann T, Leder P. Structure of the human immunoglobulin mu locus: characterization of embryonic and rearranged J and D genes.Cell. 1981;27:583–591.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Tomita A, Buchholz DR, Shi YB. Recruitment of N-CoR/SMRT- TBLR1 corepressor complex by unliganded thyroid hormone receptor for gene repression during frog development.Mol Cell Biol. 2004;24:3337–3346.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Tomita A, Buchholz DR, Obata K, Shi YB. Fusion protein of retinoic acid receptor α with promyelocytic leukemia protein or promyelocytic leukemia zinc finger protein recruits N-CoR-TBLR1 corepressor complex to repress transcriptionin vivo.J Biol Chem. 2003;278:30788–30795.

    Article  PubMed  CAS  Google Scholar 

  22. Crescenzi M, Seto M, Herzig GP, Weiss PD, Griffith RC, Korsmeyer SJ. Thermostable DNA polymerase chain amplification of t(14;18) chromosome breakpoints and detection of minimal residual disease.Proc Natl Acad Sci U S A. 1988;85:4869–4873.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Kosugi H, Towatari M, Hatano S, et al. Histone deacetylase inhibitors are the potent inducer/enhancer of differentiation in acute myeloid leukemia: a new approach to anti-leukemia therapy.Leukemia. 1999;13:1316–1324.

    Article  PubMed  CAS  Google Scholar 

  24. Hsia SC, Tomita A, Obata K, Paul B, Buchholz D, Shi YB. Role of chromatin disruption and histone acetylation in thyroid hormone receptor action: implications in the regulation of HIV-1 LTR.Histol Histopathol. 2003;18:323–331.

    PubMed  CAS  Google Scholar 

  25. Maloney DG, Grillo-Lopez AJ, White CA, et al. IDEC-C2B8 (rituximab) anti-CD20 monoclonal antibody therapy in patients with relapsed low-grade non-Hodgkin’s lymphoma.Blood. 1997;90:2188–2195.

    PubMed  CAS  Google Scholar 

  26. Maloney DG, Grillo-Lopez AJ, Bodkin DJ, et al. IDEC-C2B8: results of a phase I multiple-dose trial in patients with relapsed non-Hodgkin’s lymphoma.J Clin Oncol. 1997;15:3266–3274.

    Article  PubMed  CAS  Google Scholar 

  27. Pegoraro L, Palumbo A, Erikson J, et al. A 14;18 and an 8;14 chromosome translocation in a cell line derived from an acute B-cell leukemia.Proc Natl Acad Sci U S A. 1984;81:7166–7170.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Egger G, Liang G, Aparicio A, Jones PA. Epigenetics in human disease and prospects for epigenetic therapy.Nature. 2004;429:457–463.

    Article  PubMed  CAS  Google Scholar 

  29. Binder M, Otto F, Mertelsmann R, Veelken H, Trepel M. The epitope recognized by rituximab.Blood. 2006;108:1975–1978.

    Article  PubMed  CAS  Google Scholar 

  30. Greiner D, Bonaldi T, Eskeland R, Roemer E, Imhof A. Identification of a specific inhibitor of the histone methyltransferase SU(VAR)3-9.Nat Chem Biol. 2005;1:143–145.

    Article  CAS  Google Scholar 

  31. Kiyoi H, Naoe T. Immunoglobulin variable region structure and B-cell malignancies.Int J Hematol. 2001;73:47–53.

    Article  CAS  Google Scholar 

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Affiliations

  1. Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Tsurumai-cho 65, Showa-ku, 466-8550, Nagoya, Japan

    Akihiro Tomita, Junji Hiraga, Manabu Ninomiya, Takumi Sugimoto, Tomohiro Kinoshita & Tomoki Naoe

  2. Department of Infectious Diseases, Nagoya University School of Medicine, Nagoya, Japan

    Hitoshi Kiyoi

  3. Department of Pathology, Japanese Red Cross Nagoya First Hospital, Nagoya, Japan

    Masafumi Ito

Authors
  1. Akihiro Tomita
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  2. Junji Hiraga
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  3. Hitoshi Kiyoi
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  4. Manabu Ninomiya
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  5. Takumi Sugimoto
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  6. Masafumi Ito
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  7. Tomohiro Kinoshita
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  8. Tomoki Naoe
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Correspondence to Akihiro Tomita.

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Tomita, A., Hiraga, J., Kiyoi, H. et al. Epigenetic Regulation of CD20 Protein Expression in a Novel B-Cell Lymphoma Cell Line, RRBL1, Established from a Patient Treated Repeatedly with Rituximab-Containing Chemotherapy. Int J Hematol 86, 49–57 (2007). https://doi.org/10.1532/IJH97.07028

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  • DOI: https://doi.org/10.1532/IJH97.07028

Key words

  • CD20
  • Rituximab
  • B-cell lymphoma
  • Epigenetics