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O6-methylguanine-DNA-methyltransferase promoter methylation assessment by microdissection-assisted methylation-specific PCR and high resolution melting analysis in patients with glioblastomas

Journal of Neuro-Oncology volume 106, pages 243–250 (2012)Cite this article

Abstract

We have determined O6-methylguanine-DNA-methyltransferase (MGMT) promoter methylation status by methylation-specific polymerase chain reaction (MSP) in 22 paraffin-embedded specimens of glioblastoma multiforme. A MGMT methylation-specific high resolution melting (HRM) assay was performed to compare the methylation levels of the tumorous and non-tumorous portions of each sample, which were selectively collected using a microdissection technique. MGMT methylation was detected in 10 patients using MSP, while 8 patients had both methylated and unmethylated MGMT promoters. HRM assays showed that there was no difference in the level of methylation between tumorous and non-tumorous portions of each sample. In patients with MSP-positive tumors, the overall survival (median, 22 months) was longer as compared to those with MSP-negative tumors (median, 14 months). A correlation between the methylation status of the MGMT promoter and MGMT protein expression was observed in 12 samples. This study demonstrates that MGMT methylation is not restricted to glioblastoma cells. Additionally, methylation-specific HRM is a feasible approach that can be readily applied to the methylation analysis of MGMT. A further study will be needed to determine the dynamic change of MGMT methylation in the tumor environment.

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Acknowledgment

This study was supported by the Cancer Research Fund of the Catholic Comprehensive Cancer Institute in 2008 and the Catholic Medical Center Research Foundation made in the program year of 2009 Research Fund from the St. Vincent’s Hospital in Suwon, Korea.

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Affiliations

  1. Department of Neurosurgery, St. Vincent’s Hospital, The Catholic University of Korea, Suwon, Korea

    Seung-Ho Yang & Sang Won Lee

  2. Department of Neurosurgery, Seoul St. Mary’s Hospital, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul, 137-701, Korea

    Keun Soo Lee & Yong-Kil Hong

  3. Institute of Medical Science, St. Vincent’s Hospital, The Catholic University of Korea, Suwon, Korea

    Hea Jung Yang

  4. Asan Institute for Life Science, Seoul, Korea

    Byeong Hwan Jeon

  5. Department of Hospital Pathology, St. Mary’s Hospital, The Catholic University of Korea, Seoul, Korea

    Youn Soo Lee

  6. Department of Pathology, Microdissection Genomics Research Center, The Catholic University of Korea, Seoul, Korea

    Suk Woo Nam

  7. Department of Neurosurgery, Incheon St. Mary’s Hospital, The Catholic University of Korea, Incheon, Korea

    Dong-Sup Chung

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  1. Seung-Ho Yang
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  2. Keun Soo Lee
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Correspondence to Yong-Kil Hong.

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Yang, SH., Lee, K.S., Yang, H.J. et al. O6-methylguanine-DNA-methyltransferase promoter methylation assessment by microdissection-assisted methylation-specific PCR and high resolution melting analysis in patients with glioblastomas. J Neurooncol 106, 243–250 (2012). https://doi.org/10.1007/s11060-011-0668-4

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Keywords

  • MGMT promoter methylation
  • Microdissection
  • High resolution melting
  • MGMT protein
  • Glioblastoma
  • Polymerase chain reaction