MYC and BCL2 Protein Expression in Patients with Diffuse Large B-cell Lymphoma

Anna Evgen’evna Misyurina, A.M. Kovrigina, E.A. Baryakh, V.A. Misyurin, S.K. Kravchenko, S.M. Kulikov, E.G. Gemdzhyan, T.N. Obukhova, A.N. Kopylov, A.U. Magomedova, A.I. Vorob’ev,

DOI:

https://doi.org/10.21320/2500-2139-2015-8-1-44-53

Objective. The objective of this study is to analyze the incidence and the role of MYC and BCL2 protein expression in patients with diffuse large B-cell lymphoma (DLBCL) and to compare data of histological, immunohistochemical, genetic, and moleculobiological analyses with clinical characteristics.

Methods. 62 patients with confirmed diffuse large B-cell lymphoma (DLBCL) were enrolled in the study; they underwent treatment according to the original modified protocol NHL-BFM-90 ± R in the Hematology Research Center under the Ministry of Health of the Russian Federation. A reference group consisted of 13 DLBCL patients who underwent СНОР-like ± R therapy. In all observations, histological and immunohistochemical tests were performed in archived biopsy samples of a tumor tissue or a lymph node (paraffin blocks) using BCL2 (clone 124, Dako) and MYC antibodies (clone Y69, Epitomics). Based on C.P. Hans’s algorithm (2004), GCB and non-GCB immunohistochemical subtypes of DLBCL were determined. Standard cytogenetic tests (n = 22) and FISH analysis (n = 52) were performed in this study in order to determine locus translocation of с-MYC gene, IgH gene, t(8;14)(q24;q32), BCL2 gene, t(14;18)(q32;q21). Quantitative RT-PCR on paraffin tumor/node biopsy samples was also performed to evaluate the number of mRNAs of с-MYC and BCL2 genes.

Results. MYC expression was found in 24 (39 %) of 62 DLBCL patients and BCL2 in 36 (58 %) of 62 patients (threshold values were 40 % and 50 % of tumor cells, respectively). MYC/BCL2 coexpression was diagnosed in 15 (24 %) of 62 DLBCL patients. 4 (27 %) of 15 patients with MYC/BCL2 coexpression had a GCB-subtype of DLBCL, 73 % patients with MYC/BCL2 coexpression were diagnosed with non-GCB subtype of DLBCL (p < 0.02). c-MYC rearrangement was found in two cases (3 %), one of them had a MYC protein expression score more than 40 %. 10 (19 %) patients had one or more additional signal from 8q24 locus of c-MYC gene. We didn’t find any correlation between the presence of additional signals from c-MYC gene locus and the level of immunohistochemical expression of MYC protein ³ 40 % (p < 0.05). BCL2 gene rearrangement was detected in one case; it was accompanied by BCL2 protein immunohistochemical expression ³ 50 %. BCL2 amplification was observed in 17 (40 %) patients. There was a correlation between the amplification of BCL2 gene and immunohistochemical expression of BCL2 protein (threshold value was ³ 50 % of positive cells) (p = 0.0053). There was a direct correlation between the amount of mRNAs and MYC protein (correlation coefficient 0.86; p < 0.0001). There was no correlation between the level of BCL2 gene expression and the amount of protein (correlation coefficient 0.14; p = 0.57). Four-year overall survival for DLBCL patients treated with m-NHL-BFM-90 ± R was 71 % (in patients without MYC/BCL2 coexpression) vs 57 % (in patients with MYC/BCL2 coexpression) (p = 0.39). The probability of relapse or progression of DLBCL was significantly higher in patients with MYC/BCL2 coexpression, than in patients without MYC/BCL2 coexpression (65 % vs 15 %; p = 0.0029).

Conclusions. The MYC/BCL2 coexpression is observed mainly in DLBCL patients with non-GCB immunohistochemical subtype of the disease. The MYC/BCL2 coexpression is vital for prediction of the risk of relapses/progression of the disease in patients receiving an intensive chemotherapy according to the m-NHL-BFM-90 ± R protocol. Taking into account a relatively stable structure of the protein substrate, the obtained data may become a basis for development of a diagnostic immunohistochemical algorithm for stratification of DLBCL patients.

  • Anna Evgen’evna Misyurina Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • A.M. Kovrigina Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • E.A. Baryakh Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • V.A. Misyurin N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478 ; ФГБНУ «Российский онкологический научный центр им. Н.Н. Блохина», Каширское ш., д. 24, Москва, Российская Федерация, 115478
  • S.K. Kravchenko Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • S.M. Kulikov Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • E.G. Gemdzhyan Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • T.N. Obukhova Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • A.N. Kopylov N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478 ; ФГБНУ «Российский онкологический научный центр им. Н.Н. Блохина», Каширское ш., д. 24, Москва, Российская Федерация, 115478
  • A.U. Magomedova Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  • A.I. Vorob’ev Hematology Research Center under the Ministry of Health of the Russian Federation, 4а Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167 ; ФГБУ «Гематологический научный центр» МЗ РФ, Новый Зыковский пр-д., д. 4а, Москва, Российская Федерация, 125167
  1. Swerdlow SH, Campo E, Harris NL, et al, eds. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th edition. Lyon: IARC Press; 2008.
  2. A predictive model for aggressive non-Hodgkin’s lymphoma. The International Non-Hodgkin’s Lymphoma Prognostic Factors Project. N Engl J Med. 1993;329(14):987–94. DOI: https://doi.org/10.1056/NEJM199309303291402
  3. Frick M, Dorken B, Lenz G. New insights into the biology of molecular subtypes of diffuse large B-cell lymphoma and Burkitt lymphoma. Best Pract Res Clin Haematol. 2012;25(1):3–12. doi: 10.1016/j.beha.2012.01.003. DOI: https://doi.org/10.1016/j.beha.2012.01.003
  4. Alizadeh AA, Eisen MB, Davis RE, et al. Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling. Nature. 2000;403(6769):503–11. doi: 10.1038/35000501. DOI: https://doi.org/10.1038/35000501
  5. Rosenwald A, Wright G, Chan WC, et al. The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma. N Engl J Med. 2002;346(25):1937–47. doi: 10.1056/nejmoa012914. DOI: https://doi.org/10.1056/NEJMe020050
  6. Shipp MA, Ross KN, Tamayo P, et al. Diffuse large B-cell lymphoma outcome prediction by gene-expression profiling and supervised machine learning. Nat Med. 2002;8(1):68–74. doi: 10.1038/nm0102-68. DOI: https://doi.org/10.1038/nm0102-68
  7. Lenz G, Wright G, Dave SS, et al. Stromal gene signatures in large-B-cell lymphomas. N Engl J Med. 2008;359(22):2313–23. doi: 10.1056/nejmoa0802885. DOI: https://doi.org/10.1056/NEJMoa0802885
  8. Rosenwald A, Wright G, Leroy K, et al. Molecular diagnosis of primary mediastinal B cell lymphoma identifies a clinically favorable subgroup of diffuse large B cell lymphoma related to Hodgkin lymphoma. J Exp Med. 2003;198(6):851–62. doi: 10.1084/jem.20031074. DOI: https://doi.org/10.1084/jem.20031074
  9. Savage KJ, Monti S, Kutok JL, et al. The molecular signature of mediastinal large B-cell lymphoma differs from that of other diffuse large B-cell lymphomas and shares features with classical Hodgkin lymphoma. Blood. 2003;102(12):3871–9. doi: 10.1007/s00795-013-0038-8. DOI: https://doi.org/10.1182/blood-2003-06-1841
  10. Wright G, Tan B, Rosenwald A, et al. A gene expression-based method to diagnose clinically distinct subgroups of diffuse large B cell lymphoma. Proc Natl Acad Sci USA. 2003;100(17):9991–6. doi: 10.1073/pnas.1732008100. DOI: https://doi.org/10.1073/pnas.1732008100
  11. Мисюрина А.Е., Мисюрин В.А., Барях Е.А и др. Роль экспрессии c-MYC, BCL-2, BCL-6 в патогенезе диффузной В-крупноклеточной лимфомы. Клиническая онкогематология. 2014;7(4):512–21. DOI: https://doi.org/10.21320/2500-2139-2014-7-4-512-521
  12. [Misyurina AE, Misyurin VA, Baryakh EA, et al. Role of c-MYC, BCL-2, and BCL-6 expression in pathogenesis of diffuse large B-cell lymphoma. Klinicheskaya onkogematologiya. 2014;7(4):512–21. (In Russ)]
  13. Savage KJ, Johnson NA, Ben-Neriah S, et al. MYC gene rearrangements are associated with a poor prognosis in diffuse large B-cell lymphoma patients treated with R-CHOP chemotherapy. Blood. 2009;114(17):3533–7. doi: 10.1182/blood-2009-05-220095. DOI: https://doi.org/10.1182/blood-2009-05-220095
  14. Horn H, Ziepert M, Becher C, et al. MYC status in concert with BCL2 and BCL6 expression predicts outcome in diffuse large B-cell lymphoma. Blood. 2013;121(12):2253–63. doi: 10.1182/blood-2012-06-435842. DOI: https://doi.org/10.1182/blood-2012-06-435842
  15. Barrans S, Crouch S, Smith A, et al. Rearrangement of MYC is associated with poor prognosis in patients with diffuse large B-cell lymphoma treated in the era of rituximab. J Clin Oncol. 2010;28(20):3360–5. doi: 10.1200/jco.2009.26.3947. DOI: https://doi.org/10.1200/JCO.2009.26.3947
  16. Iqbal J, Sanger WG, Horsman DE, et al. BCL2 translocation defines a unique tumor subset within the germinal center B-cell-like diffuse large B-cell lymphoma. Am J Pathol. 2004;165(1):159–66. doi: 10.1016/s0002-9440(10)63284-1. DOI: https://doi.org/10.1016/S0002-9440(10)63284-1
  17. Ohno H, Fukuhara S. Significance of rearrangement of the BCL6 gene in B-cell lymphoid neoplasms. Leuk Lymphoma. 1997;27(1–2):53–63. doi: 10.3109/10428199709068271. DOI: https://doi.org/10.3109/10428199709068271
  18. Willis TG, Dyer MJ. The role of immunoglobulin translocations in the pathogenesis of B-cell malignancies. Blood. 2000;96(3):808–22. DOI: https://doi.org/10.1182/blood.V96.3.808
  19. Hu S, Xu-Monette ZY, Tzankov A, et al. MYC/BCL2 protein co-expression contributes to the inferior survival of activated B-cell subtype of diffuse large B-cell lymphoma and demonstrates high-risk gene expression signatures: a report from The International DLBCL Rituximab-CHOP Consortium Program Study. Blood. 2013;121(20):4021–31. doi: 10.1182/blood-2012-10-460063. DOI: https://doi.org/10.1182/blood-2012-10-460063
  20. Green TM, Young KH, Visco C, et al. Immunohistochemical Double-Hit Score Is a Strong Predictor of Outcome in Patients With Diffuse Large B-Cell Lymphoma Treated With Rituximab Plus Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone. J Clin Oncol. 2012;30(28):3460–7. doi: 10.1200/jco.2011.41.4342. DOI: https://doi.org/10.1200/JCO.2011.41.4342
  21. Johnson NA, Slack GW, Savage K, et al. Concurrent Expression of MYC and BCL2 in Diffuse Large B-Cell Lymphoma Treated With Rituximab Plus Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone. J Clin Oncol. 2012;30(28):3452–9. doi: 10.1200/jco.2011.41.0985. DOI: https://doi.org/10.1200/JCO.2011.41.0985
  22. Valera A, Lopez-Guillermo A, Cardesa-Salzmann T. MYC protein expression and genetic alterations have prognostic impact in patients with diffuse large B-cell lymphoma treated with immunochemotherapy. Haematologica. 2013;98(10):1554–62. doi: 10.3324/haematol.2013.086173. DOI: https://doi.org/10.3324/haematol.2013.086173
  23. Магомедова А.У., Кравченко С.К., Кременецкая AM. и др. Модифицированная программа NHL-BFM-90 в лечении больных диффузной В-крупноклеточной лимфосаркомой. Терапевтический архив. 2006;10:44–7.
  24. [Magomedova AU, Kravchenko SK, Kremenetskaya AM, et al. Modified NHL-BFM-90 protocol in treatment of diffuse large B-cell lymphosarcoma. Terapevticheskii arkhiv. 2006;10:44–7. (In Russ)]
  25. Hans CP, Weisenburger DD, Greiner TC, Gascoyne RD. Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray. Blood. 2004;103(1):275–82. doi: 10.1182/blood-2003-05-1545. DOI: https://doi.org/10.1182/blood-2003-05-1545
  26. Green TM, Nielsen O, de Stricker K, et al. High levels of nuclear MYC protein predict the presence of MYC rearrangement in diffuse large B-cell lymphoma. Am J Surg Pathol. 2012;36(4):612–9. doi: 10.1097/pas.0b013e318244e2ba. DOI: https://doi.org/10.1097/PAS.0b013e318244e2ba
  27. Cook JR, Goldman B, Tubbs RR. Clinical significance of MYC expression and/or “high-grade” morphology in non-Burkitt, diffuse aggressive B-cell lymphomas: a SWOG S9704 correlative study. Am J Surg Pathol. 2014;38(4):494–501. doi: 10.1097/PAS.0000000000000147. DOI: https://doi.org/10.1097/PAS.0000000000000147
  28. Leucci E, Cocco M, Onnis A, et al. MYC translocation-negative classical Burkitt lymphoma cases: an alternative pathogenetic mechanism involving miRNA deregulation. J Pathol. 2008;216(4):440–50. doi: 10.1002/path.2410. DOI: https://doi.org/10.1002/path.2410
  29. Onnis A, De Falco G, Antonicelli G, et al. Аlteration of microRNAs regulated by c-MYC in Burkitt lymphoma. PLoS One. 2010;5(9);e12960. doi: 10.1371/journal.pone.0012960. DOI: https://doi.org/10.1371/journal.pone.0012960
  30. Kluin PM. Origin And Migration of Follicular Lymphoma Cells. Haematologica. 2013;98(9):1331–3. doi: 10.3324/haematol.2013.091546. DOI: https://doi.org/10.3324/haematol.2013.091546
  31. Ameres SL, Zamore PD. Diversifying microRNA sequence and function. Nat Rev Mol Cell Biol. 2013;14(8):475–88. doi: 10.1038/nrm3611. DOI: https://doi.org/10.1038/nrm3611
  32. Мангасарова Я.К., Мисюрин А.В., Магомедова А.У. и др. Молекулярная диагностика первичной медиастинальной В-клеточной лимфомы и диффузной В-крупноклеточной лимфомы с первичным вовлечением лимфоузлов средостения. Клиническая онкогематология. 2011;4(2):142–5.
  33. [Mangasarova YaK, Misyurin AV, Magomedova AU, et al. Molecular diagnostics of primary mediastinal B-cell lymphoma and diffuse large B-cell lymphoma with primary involvement of mediastinal lymph nodes. Klinicheskaya onkogematologiya. 2011;4(2):142–5. (In Russ)]
  34. Liu Y, Hernandez AM, Shibata D, Cortopassi GA. BCL2 translocation frequency rises with age in humans. Proc Natl Acad Sci USA. 1994;91(19):8910–4. doi: 10.1073/pnas.91.19.8910. DOI: https://doi.org/10.1073/pnas.91.19.8910
  35. Harries LW, Hernandez D, Henley W. Human aging is characterized by focused changes in gene expression and deregulation of alternative splicing. Aging Cell. 2011;10(5):868–78. doi: 10.1111/j.1474-9726.2011.00726.x. DOI: https://doi.org/10.1111/j.1474-9726.2011.00726.x
  36. Dunleavy K, Pittaluga S, Shovlin M. Concurrent Expression Of MYC/BCL2 Protein In Newly Diagnosed DLBCL Is Not Associated With An Inferior Survival Following EPOCH-R Therapy. Blood. 2013;122(21):3029. DOI: https://doi.org/10.1182/blood.V122.21.3029.3029

Keywords:

DLBCL, intensive therapy, MYC/BCL2 coexpression, immunohistochemistry, unfavorable prognostic factor

License

Copyright (c) 2015 Clinical Oncohematology

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Downloads

Download data is not yet available.
2015-1

Downloads

Published

01.01.2015

Issue

Vol. 8 No. 1 (2015)
LYMPHOID MALIGNANCIES

How to Cite

Misyurina A.E., Kovrigina A.M., Baryakh E.A., et al. MYC and BCL2 Protein Expression in Patients with Diffuse Large B-cell Lymphoma. Clinical Oncohematology. 2015;8(1):44–53. doi:10.21320/2500-2139-2015-8-1-44-53.

Most read articles by the same author(s)

1 2 3 4 5 6 7 8 > >>