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International Journal of Hematology

, Volume 83, Issue 4, pp 341–347 | Cite as

Loss of O6-Methylguanine-DNA Methyltransferase Protein Expression Is a Favorable Prognostic Marker in Diffuse Large B-Cell Lymphoma

  • Toshihito Ohno
  • Junji Hiraga
  • Haruhiko Ohashi
  • Chiho Sugisaki
  • Eika Li
  • Haruhiko Asano
  • Tastuya Ito
  • Hirokazu Nagai
  • Yoriko Yamashita
  • Naoyoshi Mori
  • Tomohiro Kinoshita
  • Tomoki Naoe
Case Report

Abstract

Although aberrant promoter hypermethylation of O6-methylguanine-DNA methyltransferase (MGMT) is a favorable prognostic marker in patients with diffuse large B-cell lymphoma (DLBCL), MGMT protein expression has not been thoroughly examined. The aim of this study was to evaluate the clinical implication of MGMT protein expression and its correlation with promoter hypermethylation of the gene. We investigated MGMT protein expression by immunohistochemical analysis of 63 DLBCL patients who received cyclophosphamide as part of multidrug regimens. In addition, promoter methylation of the MGMT gene was analyzed by a methylation-specific polymerase chain reaction assay, and correlations with chemotherapeutic effect and prognosis were statistically evaluated. Immunohistochemical assay results for MGMT protein were negative in 30.2% of patients with newly diagnosed DLBCL. Immunostaining results were closely correlated with the methylation status of the promoter. Promoter DNA methylation of the gene was not detected in 34 (81.0%) of 42 tumor samples determined to be MGMT-positive DLBCL by immunostaining and was detected in 15 (88.2%) of 17 cases of MGMT-negative DLBCL. Overall survival (OS) and disease-free survival (DFS) rates were significantly higher in MGMT-negative patients than in MGMT-positive patients (5-year OS, 81.3% versus 56.6% [P = .0375]; 5-year DFS, 66.3% versus 39.9% [P = .0121]). The combined rate for complete response (CR) plus unconfirmed CR was significantly higher in MGMT-negative patients (15/19, 79.0%) than in MGMT-positive patients (25/44, 56.8%) (P = .0488). A multivariate analysis showed that absence of MGMT expression was an independent prognostic factor for OS (relative risk, 4.09; P = .0258). Lack of MGMT protein expression is associated with aberrant promoter DNA methylation and appears to be a useful marker for predicting the survival of DLBCL patients.

Key words

MGMT Hypermethylation Diffuse large B-cell lymphoma Expression 

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References

  1. 1.
    Pegg AE, Dolan ME, Moschel RC. Structure, function, and inhibition of O 6-alkylguanine-DNA alkyltransferase. Prog Nucleic Acid Res Mol Biol. 1995;51:167–223.CrossRefPubMedGoogle Scholar
  2. 2.
    Silber JR, Mueller BA, Ewers TG, Berger MS. Comparison of O 6-methylguanine-DNA methyltransferase activity in brain tumors and adjacent normal brain. Cancer Res. 1993;53:3416–3420.PubMedGoogle Scholar
  3. 3.
    Belanich M, Pastor M, Randall T, et al. Retrospective study of the correlation between the DNA repair protein alkyltransferase and survival of brain tumor patients treated with carmustine. Cancer Res. 1996;56:783–788.PubMedGoogle Scholar
  4. 4.
    Mattern J, Eichhorn U, Kaina B, Volm M. O 6-methylguanine-DNA methyltransferase activity and sensitivity to cyclophosphamide and cisplatin in human lung tumor xenografts. Int J Cancer. 1998;77:919–922.Google Scholar
  5. 5.
    Silber JR, Bobola MS, Ghatan S, Blank A, Kolstoe DD, Berger MS. O 6-methylguanine-DNA methyltransferase activity in adult gliomas:relation to patient and tumor characteristics. Cancer Res. 1998;58:1068–1073.Google Scholar
  6. 6.
    Jaeckle KA, Eyre HJ, Townsend JJ, et al. Correlation of tumor O 6 methylguanine-DNA methyltransferase levels with survival of malignant astrocytoma patients treated with bis-chloroethylnitrosourea: a Southwest Oncology Group study. J Clin Oncol. 1998;16:3310–3315.CrossRefPubMedGoogle Scholar
  7. 7.
    Silber JR, Blank A, Bobola MS, Ghatan S, Kolstoe DD, Berger MS. O 6-methylguanine-DNA methyltransferase-deficient phenotype in human gliomas: frequency and time to tumor progression after alkylating agent-based chemotherapy. Clin Cancer Res. 1999;5:807–814.Google Scholar
  8. 8.
    Pegg AE, Byers TL. Repair of DNA containing O 6-alkylguanine. FASEB J. 1992;6:2302–2310.CrossRefPubMedGoogle Scholar
  9. 9.
    Watts GS, Pieper RO, Costello JF, Peng YM, Dalton WS, Futscher BW. Methylation of discrete regions of the O 6-methylguanine DNA methyltransferase (MGMT) CpG island is associated with hete-rochromatinization of the MGMT transcription start site and silencing of the gene. Mol Cell Biol. 1997;17:5612–5619.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Qian XC, Brent TP. Methylation hot spots in the 5′ flanking region denote silencing of the O 6-methylguanine-DNA methyltransferase gene. Cancer Res. 1997;57:3672–3677.PubMedGoogle Scholar
  11. 11.
    Danam RP, Qian XC, Howell SR, Brent TP. Methylation of selected CpGs in the human O 6-methylguanine-DNA methyltransferase promoter region as a marker of gene silencing. Mol Carcinog. 1999;24:85–89.CrossRefPubMedGoogle Scholar
  12. 12.
    Esteller M, Hamilton SR, Burger PC, Baylin SB, Herman JG Inactivation of the DNA repair gene O 6-methylguanine-DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia. Cancer Res. 1999;59:793–797.PubMedGoogle Scholar
  13. 13.
    Herfarth KK, Brent TP, Danam RP, et al. A specific CpG methylation pattern of the MGMT promoter region associated with reduced MGMT expression in primary colorectal cancers. Mol Carcinog. 1999;24:90–98.CrossRefPubMedGoogle Scholar
  14. 14.
    Yin D, Xie D, Hofmann WK, et al. DNA repair gene O 6-methylguanine-DNA methyltransferase: promoter hypermethylation associated with decreased expression and G:C to AT mutations of p53 in brain tumors. Mol Carcinog. 2003;36:23–31.CrossRefPubMedGoogle Scholar
  15. 15.
    Esteller M, Toyota M, Sanchez-Cespedes M, et al. Inactivation of the DNA repair gene O 6-methylguanine-DNA methyltransferase by promoter hypermethylation is associated with G to A mutations in K-ras in colorectal tumorigenesis. Cancer Res. 2000;60:2368–2371.PubMedGoogle Scholar
  16. 16.
    Esteller M, Risques RA,Toyota M, et al. Promoter hypermethylation of the DNA repair gene O 6-methylguanine-DNA methyltransferase is associated with the presence of G:C to A:T transition mutations in p53 in human colorectal tumorigenesis. Cancer Res. 2001;61:4689–4692.PubMedGoogle Scholar
  17. 17.
    Wolf P, Hu YC, Doffek K, Sidransky D,Ahrendt SA. O 6-Methylguanine-DNA methyltransferase promoter hypermethylation shifts the p53 mutational spectrum in non-small cell lung cancer. Cancer Res. 2001;61:8113–8117.Google Scholar
  18. 18.
    Nakamura M, Watanabe T, Yonekawa Y, Kleihues P, Ohgaki H. Promoter methylation of the DNA repair gene MGMT in astrocytomas is frequently associated with G:C → AT mutations of the TP53 tumor suppressor gene. Carcinogenesis. 2001;22:1715–1719.CrossRefPubMedGoogle Scholar
  19. 19.
    Esteller M, Gaidano G, Goodman SN, et al. Hypermethylation of the DNA repair gene O 6-methylguanine DNA methyltransferase and survival of patients with diffuse large B-cell lymphoma. J Natl Cancer Inst. 2002;94:26–32.CrossRefPubMedGoogle Scholar
  20. 20.
    Esteller M, Garcia-Foncillas J, Andion E, et al. Inactivation of the DNA-repair gene MGMT and the clinical response of gliomas to alkylating agents. N Engl J Med. 2000;343:1350–1354.CrossRefPubMedGoogle Scholar
  21. 21.
    Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci U S A. 1996;93:9821–9826.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Brent TP, von Wronski M, Pegram CN, Bigner DD. Immunoaffinity purification of human O 6-alkylguanine-DNA alkyltransferase using newly developed monoclonal antibodies. Cancer Res. 1990;50:58–61.PubMedGoogle Scholar
  23. 23.
    Harris NL, Jaffe ES, Stein H, et al. A revised European-American classification of lymphoid neoplasms: a proposal from the International Lymphoma Study Group. Blood. 1994;84:1361–1392.PubMedGoogle Scholar
  24. 24.
    Carbone PP, Kaplan HS, Musshoff K, Smithers DW, Tubiana M. Report of the Committee on Hodgkin’s Disease Staging Classification. Cancer Res. 1971;31:1860–1861.PubMedGoogle Scholar
  25. 25.
    Lister TA, Crowther D, Sutcliffe SB, et al. Report of a committee convened to discuss the evaluation and staging of patients with Hodgkin’s disease: Cotswolds meeting. J Clin Oncol. 1989;7:1630–1636.CrossRefPubMedGoogle Scholar
  26. 26.
    Mori M, Kitamura K, Masuda M, et al. Long-term results of a multicenter randomized, comparative trial of modified CHOP versus THP-COP versus THP-COPE regimens in elderly patients with non-Hodgkin’s lymphoma. Int J Hematol. 2005;81:246–254.CrossRefPubMedGoogle Scholar
  27. 27.
    Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981;29:577–580.CrossRefPubMedGoogle Scholar
  28. 28.
    Hashimoto M, Yamashita Y, Mori N. Immunohistochemical detection of CD79a expression in precursor T cell lymphoblastic lymphoma/leukaemias. J Pathol. 2002;197:341–347.CrossRefPubMedGoogle Scholar
  29. 29.
    Pinyol M, Hernandez L, Cazorla M, et al. Deletions and loss of expression of P16INK4a and P21Waf1 genes are associated with aggressive variants of mantle cell lymphomas. Blood. 1997;89:272–280.PubMedGoogle Scholar
  30. 30.
    Kees UR, Burton PR, Lu C, Baker DL. Homozygous deletion of the p16/MTS1 gene in pediatric acute lymphoblastic leukemia is associated with unfavorable clinical outcome. Blood. 1997;89:4161–4166.PubMedGoogle Scholar
  31. 31.
    Wong IH, Ng MH, Huang DP, Lee JC Aberrant p15 promoter methylation in adult and childhood acute leukemias of nearly all morphologic subtypes: potential prognostic implications. Blood. 2000;95:1942–1949.PubMedGoogle Scholar
  32. 32.
    Sanchez-Beato M, Saez AI, Navas IC, et al. Overall survival in aggressive B-cell lymphomas is dependent on the accumulation of alterations in p53, p16, and p27. Am J Pathol. 2001;159:205–213.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Graf-Einsiedel H, Taube T, Hartmann R, et al. Prognostic value of p16 INK4a gene deletions in pediatric acute lymphoblastic leukemia. Blood. 2001;97:4002–4004.CrossRefPubMedGoogle Scholar
  34. 34.
    Roman-Gomez J, Castillejo JA, Jimenez A, et al. 5′ CpG island hypermethylation is associated with transcriptional silencing of the p21cip1/Waf1/sdi1 gene and confers poor prognosis in acute lymphoblastic leukemia. Blood. 2002;99:2291–2296.CrossRefPubMedGoogle Scholar
  35. 35.
    Uchida T, Kinoshita T, Nagai H, et al. Hypermethylation of the p15 INK4B gene in myelodysplastic syndromes. Blood. 1997;90:1403–1409.PubMedGoogle Scholar
  36. 36.
    Kawano S, Miller CW, Gombart AF, et al. Loss of p73 gene expression in leukemias/lymphomas due to hypermethylation. Blood. 1999;94:1113–1120.PubMedGoogle Scholar
  37. 37.
    Katzenellenbogen RA, Baylin SB, Herman JG. Hypermethylation of the DAP-kinase CpG island is a common alteration in B-cell malignancies. Blood. 1999;93:4347–4353.PubMedGoogle Scholar
  38. 38.
    Singal R, Ginder GD. DNA methylation. Blood. 1999;93:4059–4070.PubMedGoogle Scholar
  39. 39.
    Toyota M, Kopecky KJ, Toyota MO, Jair KW, Willman CL, Issa JP Methylation profiling in acute myeloid leukemia. Blood. 2001;97:2823–2829.CrossRefPubMedGoogle Scholar
  40. 40.
    Roman J, Castillejo JA, Jimenez A, et al. Hypermethylation of the calcitonin gene in acute lymphoblastic leukaemia is associated with unfavourable clinical outcome. Br J Haematol. 2001;113:329–338.CrossRefPubMedGoogle Scholar
  41. 41.
    Kinoshita T. Epigenetic inactivation of tumor suppressor genes in hematologic malignancies. Int J Hematol. 2004;80:108–119.CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society of Hematology 2006

Authors and Affiliations

  • Toshihito Ohno
    • 1
  • Junji Hiraga
    • 1
  • Haruhiko Ohashi
    • 3
  • Chiho Sugisaki
    • 1
  • Eika Li
    • 1
  • Haruhiko Asano
    • 1
  • Tastuya Ito
    • 1
  • Hirokazu Nagai
    • 3
  • Yoriko Yamashita
    • 2
  • Naoyoshi Mori
    • 2
  • Tomohiro Kinoshita
    • 1
  • Tomoki Naoe
    • 1
  1. 1.Departments of Hematology and OncologyNagoya University Graduate School of MedicineJapan
  2. 2.Departments of Pathology of Biological ResponseNagoya University Graduate School of MedicineJapan
  3. 3.Department of Clinical ResearchNagoya National HospitalNagoyaJapan

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