Careful exclusion of non-neoplastic brain components is required for an appropriate evaluation of O6-methylguanine-DNA methyltransferase status in glioma: Relationship between immunohistochemistry and methylation analysis

Ken Sasai, Miho Nodagashira, Hiroshi Nishihara, Eiko Aoyanagi, Lei Wang, Masahito Katoh, Junichi Murata, Yoshimaru Ozaki, Tamio Ito, Shin Fujimoto, Sadao Kaneko, Kazuo Nagashima, Shinya Tanaka

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Evaluation of O-methylguanine-DNA methyltransferase (MGMT) expression is important for antiglioma therapy as many clinical trials have demonstrated that promoter hypermethylation and low level expression of MGMT are associated with an enhanced response to alkylating agents. However, here we report that the current strategies used to evaluate MGMT status in gliomas are unreliable. We observed discordance in the MGMT expression status when immunohistochemical evaluation and polymerase chain reaction-based methylation assessments were used: 73% of gliomas with methylated MGMT promoter had substantial numbers of MGMT-immunopositive tumor cells. Furthermore, when MGMT expression was tested in tumor homogenates using reverse transcription-polymerase chain reaction, 43% of tumors were found positive, in comparison to only 24%, when histologic samples were assayed immunohistochemically. To explain these inconsistencies we undertook a detailed immunohistochemical evaluation of tumor samples and found that some gliomas demonstrated remarkably high expression of MGMT in the entire tumor whereas others contained only a small immunopositive area. Additionally, we found that gliomas contained various types of non-neoplastic cells expressing MGMT, including lymphocytes, vascular endothelial cells, and macrophages/microglias, which contribute to overall MGMT expression detected in tumor homogenates, and thus result in overestimation of tumor MGMT expression. Therefore, to correctly establish MGMT expression in the tumor, which could be informative of glioma sensitivity to alkylating agents, exclusion of non-neoplastic brain components from analysis is required.

Original languageEnglish
Pages (from-to)1220-1227
Number of pages8
JournalAmerican Journal of Surgical Pathology
Volume32
Issue number8
DOIs
Publication statusPublished - 2008 Aug 1
Externally publishedYes

Fingerprint

Methyltransferases
Glioma
Methylation
Immunohistochemistry
DNA
Brain
Neoplasms
Alkylating Agents
O-(6)-methylguanine
Polymerase Chain Reaction
Reverse Transcription
Endothelial Cells
Macrophages
Clinical Trials
Lymphocytes

Keywords

  • CD68
  • Glioma
  • MGMT
  • Promoter methylation

ASJC Scopus subject areas

  • Anatomy
  • Surgery
  • Pathology and Forensic Medicine

Cite this

Careful exclusion of non-neoplastic brain components is required for an appropriate evaluation of O6-methylguanine-DNA methyltransferase status in glioma : Relationship between immunohistochemistry and methylation analysis. / Sasai, Ken; Nodagashira, Miho; Nishihara, Hiroshi; Aoyanagi, Eiko; Wang, Lei; Katoh, Masahito; Murata, Junichi; Ozaki, Yoshimaru; Ito, Tamio; Fujimoto, Shin; Kaneko, Sadao; Nagashima, Kazuo; Tanaka, Shinya.

In: American Journal of Surgical Pathology, Vol. 32, No. 8, 01.08.2008, p. 1220-1227.

Research output: Contribution to journalArticle

Sasai, Ken ; Nodagashira, Miho ; Nishihara, Hiroshi ; Aoyanagi, Eiko ; Wang, Lei ; Katoh, Masahito ; Murata, Junichi ; Ozaki, Yoshimaru ; Ito, Tamio ; Fujimoto, Shin ; Kaneko, Sadao ; Nagashima, Kazuo ; Tanaka, Shinya. / Careful exclusion of non-neoplastic brain components is required for an appropriate evaluation of O6-methylguanine-DNA methyltransferase status in glioma : Relationship between immunohistochemistry and methylation analysis. In: American Journal of Surgical Pathology. 2008 ; Vol. 32, No. 8. pp. 1220-1227.
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