Advertisement

Journal of Neuro-Oncology

, Volume 87, Issue 1, pp 71–78 | Cite as

Variation of O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation in serial samples in glioblastoma

  • Jonathon F. Parkinson
  • Helen R. Wheeler
  • Adele Clarkson
  • Catriona A. McKenzie
  • Michael T. Biggs
  • Nicholas S. Little
  • Raymond J. Cook
  • Marinella Messina
  • Bruce G. Robinson
  • Kerrie L. McDonald
Clinical-patient studies

Abstract

Methylation of the promoter region of the O 6 -methylguanine-DNA methyltransferase (MGMT) gene is known to be predictive of response to temozolomide treatment in patients with glioblastoma. Contrastingly, little is known about variation in the methylation status of the MGMT promoter after treatment or across different regions of the same tumor. About 22 samples from 10 patients who had undergone multiple resections of a glioblastoma were examined with promoter sequencing. Of these, 20 were also analyzed using Methylation Specific PCR (MSP). The methylation status of the MGMT promoter was altered in the specimens obtained pre and post treatment in 2 of 9 samples as assessed by MSP and 7 out of 10 patients as assessed by promoter sequencing. In four patients, the MGMT promoter was unmethylated at primary surgery, but displayed some methylation (32, 44, 12, and 4%) on post-treatment sampling. Alteration in MSP status from unmethylated to methylated was also observed in 2 of these 4 patients. In another patient, methylation increased from 40% on initial sampling to 68% on the second sample. The remaining two patients initially demonstrated some degree of methylation (72% and 12%); subsequent sampling showed no methylation of the MGMT promoter. To ensure variable methylation status was not due to intra-tumoral variability, three to four specimens were sampled from different regions of large glioblastomas (n = 7). Promoter sequencing revealed minimal variation in methylation in all but two sites examined. Immunohistochemistry also demonstrated minimal change in MGMT expression across the tumors. This suggests that variation in MGMT promoter methylation can occur within the same tumor after treatment, necessitating caution in clinical decision-making based on this analysis.

Keywords

Glioma MGMT Temozolomide Methylation DNA repair 

Notes

Acknowledgment

We would like to acknowledge the contribution of Dr Anthony Gill, Department of Anatomical Pathology, Pacific Laboratory Medical Services, Royal North Shore Hospital to the optimisation of the immunohistochemistry.

References

  1. 1.
    Kleihues P, Burger PC, Aldape KD, Brat DJ, Biernat W, Bigner DD, Nakazato Y, Plate KH, Giangaspero F, von Deimling A, Ohgaki H, Cavenee WK (2007) Glioblastoma. In: Louis DN, Ohgaki H, Wiestler OD, Cavenee WK (eds) WHO classification of tumours of the central nervous system. International Agency for Research on Cancer, Lyon, pp 33–49Google Scholar
  2. 2.
    Walker MD, Alexander EJ, WE H, MacCarty CS, Mahaley MSJ, Mealey JJ (1978) Evaluation of BCNU and/or radiotherapy in the treatment of anaplastic gliomas. A cooperative clinical trial. J Neurosurg 49:333–343PubMedCrossRefGoogle Scholar
  3. 3.
    Stewart LA (2002) Chemotherapy in adult high-grade glioma: a systematic review and meta-analysis of individual patient data from 12 randomised trials. Lancet 359:1011–1018PubMedCrossRefGoogle Scholar
  4. 4.
    Stupp R, Mason WP, Van Den Bent MJ, Weller M, Fisher B, Taphoorn M, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO (2005) Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med 352:987–996PubMedCrossRefGoogle Scholar
  5. 5.
    Hegi ME, Diserens A-C, Gorlia T, Hamou M-F, De Tribolet N, Weller M, Kros JM, Hainfellner JA, Mason W, Mariani L, Bromberg JE, Hau P, Mirimanoff RO, Cairncross JG, Janzer RC, Stupp R (2005) MGMT silencing and benefit from temozolomide in glioblastoma. N Engl J Med 352:997–1003PubMedCrossRefGoogle Scholar
  6. 6.
    Pegg AE (1990) Mammalian O6-alkylguanine-DNA alkyltransferase: regulation and importance in response to alkylating carcinogenic and therapeutic agents. Cancer Res 50:6119–6129PubMedGoogle Scholar
  7. 7.
    Esteller M, Corn PG, Baylin SB, Herman JG (2001) A gene hypermethylation profile of human cancer. Cancer Res 61:3225–3229PubMedGoogle Scholar
  8. 8.
    Biswas T, Ramana CV, Srinivasan G, Boldogh I, Hazra TK, Chen Z, Tano K, Thompson EB, Mitra S (1999) Activation of O6-methylguanine-DNA methyltransferase gene by glucocortocoid hormone. Oncogene 18:525–532PubMedCrossRefGoogle Scholar
  9. 9.
    Gerson SL, Trey JE, Miller K, Berger NA (1986) Comparison of O6-alkylguanine-DNA alkyltransferase activity based on cellular DNA content in human, rat and mouse tissues. Carcinogenesis 7:745–749PubMedCrossRefGoogle Scholar
  10. 10.
    Esteller M, Hamilton SR, Burger PC, Baylin SB, Herman JG (1999) Inactivation of the DNA repair gene O6-methylguanine-DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia. Cancer Res 59:793–797PubMedGoogle Scholar
  11. 11.
    Esteller M, Herman JG (2004) Generating mutations but providing chemosensitivity: the role of O6-methylguanine DNA methyltransferase in human cancer. Oncogene 23:1–8PubMedCrossRefGoogle Scholar
  12. 12.
    Silber JR, Blank A, Bobola MS, Ghatan S, Kolstoe DD, Berger MS (1999) O6-methylguanine-DNA methyltransferase-deficient phenotype in human gliomas: frequency and time to tumor progression after alkylating agent-based chemotherapy. Clin Cancer Res 5:807–814PubMedGoogle Scholar
  13. 13.
    Hegi ME, Diserens A-C, Godard S, Dietrich P-Y, Regli L, Ostermann S, Otten P, Ven Melle G, De Tribolet N, Stupp R (2004) Clinical trial substantiates the predictive value of O-6 Methylguanine-DNA Methyltransferase promoter methylation in glioblastoma patients treated with temozolomide. Clin Cancer Res 10:1871–1874PubMedCrossRefGoogle Scholar
  14. 14.
    Watanabe T, Katayama Y, Komine C, Yoshino A, Ogino A, Ohto T, Fukushima T (2005) O 6 -Methylguanine-DNA Methyltransferase methylation and TP53 mutation in malignant astrocytomas and their relationships with clinical course. Int J Cancer 113:581–587PubMedCrossRefGoogle Scholar
  15. 15.
    Brell M, Tortosa A, Verger E, Gil JM, Vinolas N, Villa S, Acebes JJ, Caral L, Pujol T, Ferrer I, Ribalta T, Graus F (2005) Prognostic significance of O6-methylguanine-DNA methyltransferase determined by promoter hypermethylation and immunohistochemical expression in anaplastic gliomas. Clin Cancer Res 11:5167–5174PubMedCrossRefGoogle Scholar
  16. 16.
    Blanc JL, Wager M, Guilhot J, Kusy S, Bataille B, Chantereau T, Lapierre F, Larsen CJ, Karayan-Tapon L (2004) Correlation of clinical features and methylation status of MGMT gene promoter in glioblastomas. J Neurooncol 68:275–283PubMedCrossRefGoogle Scholar
  17. 17.
    Mollemann M, Wolter M, Felsberg J, Collins VP, Reifenberger G (2005) Frequent promoter hypermethylation and low expression of the MGMT gene in oligodendroglial tumors. Int J Cancer 113:379–385PubMedCrossRefGoogle Scholar
  18. 18.
    Grasbon-Frodl EM, Kreth FW, Ruiter M, Schnell O, Bise K, Felsberg J, Reifenberger G, Tonn JC, Kretzschmar HA (2007) Intratumoral homogeneity of MGMT promoter hypermethylation as demonstrated in serial stereotactic specimens from anaplastic astrocytomas and glioblastomas. Int J Cancer 121:2458–2464PubMedCrossRefGoogle Scholar
  19. 19.
    Bhakat KK, Mitra S (2003) CpG methylation-dependent repression of the human O6-methylguanine-DNA methyltransferase gene linked to chromatin structure alteration. Carcinogenesis 24:1337–1345PubMedCrossRefGoogle Scholar
  20. 20.
    Coons SW, Johnson PC (1993) Regional heterogeneity in the DNA content of human gliomas. Cancer 72:3052–3060PubMedCrossRefGoogle Scholar
  21. 21.
    Hassler M, Seidl S, Fazeny-Doerner B, Preusser M, Hainfellner J, Rossler K, Prayer D, Marosi C (2006) Diversity of cytogenetic and pathohistologic profiles in glioblastoma. Cancer Genet Cytogenet 166:46–55PubMedCrossRefGoogle Scholar
  22. 22.
    Coons SW, Johnson PC (1993) Regional heterogeneity in the proliferative activity of human gliomas as measured by the Ki-67 labeling index. J Neuropathol Exp Neurol 52:609–618PubMedCrossRefGoogle Scholar
  23. 23.
    Ito M, Wakabayashi T, Natsume A, Hatano H, Fujii M, Yoshida J (2007) Genetically heterogeneous glioblastoma recurring with disappearance of 1p/19q losses: case report. Neurosurgery 61:E168–169; discussion E169PubMedCrossRefGoogle Scholar
  24. 24.
    Rastetter M, Schagdarsurengin U, Lahtz C, Fiedler E, Marsch W, Dammann R, Helmbold P (2007) Frequent intra-tumoural heterogeneity of promoter hypermethylation in malignant melanoma. Histol Histopathol 22:1005–1015PubMedGoogle Scholar
  25. 25.
    Christman JK (2002) 5-Azacytidine and 5-aza-2′-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy. Oncogene 21:5483–5495PubMedCrossRefGoogle Scholar
  26. 26.
    Pogribny I, Koturbash I, Tryndyak V, Hudson D, Stevenson SM, Sedelnikova O, Bonner W, Kovalchuk O (2005) Fractionated low-dose radiation exposure leads to accumulation of DNA damage and profound alterations in DNA and histone methylation in the murine thymus. Mol Cancer Res 3:553–561PubMedCrossRefGoogle Scholar
  27. 27.
    Kovalchuk O, Burke P, Besplug J, Slovack M, Filkowski J, Pogribny I (2004) Methylation changes in muscle and liver tissues of male and female mice exposed to acute and chronic low-dose X-ray-irradiation. Mutat Res 548:75–84PubMedGoogle Scholar
  28. 28.
    Pieper RO (1997) Understanding and manipulating O6-methylguanine-DNA methyltransferase expression. Pharmacol Ther 74:285–297PubMedCrossRefGoogle Scholar
  29. 29.
    Liu L, Gerson SL (2006) Targeted modulation of MGMT: clinical implications. Clin Cancer Res 12:328–331PubMedCrossRefGoogle Scholar
  30. 30.
    Wick A, Felsberg J, Steinbach JP, Herrlinger U, Platten M, Blaschke B, Meyermann R, Reifenberger G, Weller M, Wick W (2007) Efficacy and tolerability of temozolomide in an alternating weekly regimen in patients with recurrent glioma. J Clin Oncol 25:3357–3361PubMedCrossRefGoogle Scholar
  31. 31.
    Esteller M (2006) Epigenetics provides a new generation of oncogenes and tumour-suppressor genes. Br J Cancer 94:179–183PubMedCrossRefGoogle Scholar
  32. 32.
    Clark SJ, Melki J (2002) DNA methylation and gene silencing in cancer: which is the guilty party? Oncogene 21:5380–5387PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2007

Authors and Affiliations

  • Jonathon F. Parkinson
    • 1
    • 2
  • Helen R. Wheeler
    • 1
    • 3
  • Adele Clarkson
    • 4
  • Catriona A. McKenzie
    • 4
  • Michael T. Biggs
    • 2
  • Nicholas S. Little
    • 2
  • Raymond J. Cook
    • 2
  • Marinella Messina
    • 1
  • Bruce G. Robinson
    • 1
  • Kerrie L. McDonald
    • 1
  1. 1.Cancer Genetics Group, Kolling Institute of Medical ResearchUniversity of SydneySt LeonardsAustralia
  2. 2.Department of NeurosurgeryRoyal North Shore HospitalSt LeonardsAustralia
  3. 3.Department of Medical OncologyRoyal North Shore HospitalSt LeonardsAustralia
  4. 4.Department of Anatomical Pathology, Pacific Laboratory Medicine ServicesRoyal North Shore HospitalSt LeonardsAustralia

Personalised recommendations