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Open Access 01-06-2018

MGMT assessment in pituitary adenomas: comparison of different immunohistochemistry fixation chemicals

Authors: Alexander S. G. Micko, Romana Höftberger, Adelheid Wöhrer, Matthias Millesi, Engelbert Knosp, Stefan Wolfsberger

Published in: Pituitary | Issue 3/2018

Abstract

Purpose

Despite the established role of O6-methyl-guanine-DNA methyltransferase (MGMT) as a marker for temozolomide response, consensus of the most reliable method to assess MGMT expression in pituitary adenomas is still missing. Currently, immunohistochemistry (IHC) assessment of formaldehyde fixed tissue samples is most widely used in a semiquantitative description. As formaldehyde fails to completely preserve nucleic acids, RCL2, an alcohol-based formaldehyde-free fixative, has been proposed as a more reliable alternative in terms of cell stability. Furthermore, as the current method of IHC is semiquantitative and observer-dependent, pyrosequencing, an objective tool to evaluate the methylation status of the MGMT promoter, has emerged as a reliable and accurate alternative. The aim of this study was to validate the current IHC method for assessment of MGMT protein expression in pituitary adenomas.

Methods

The tissue samples of 8 macroadenomas with positive IHC MGMT expression (> 50%) were investigated: first, we compared the time dependent stability of MGMT protein expression after pituitary adenoma removal between formaldehyde vs. RCL2. Then, we compared positive IHC MGMT expression with methylated promoter status using pyrosequencing.

Results

In the first 12 h after adenoma removal, tissue samples remained MGMT positive in significantly more samples when fixated with formaldehyde than with RCL2, respectively (96 vs. 81%, p = 0.025).

Conclusion

Our data confirm that the current method using formaldehyde tissue fixation and IHC reveals stable and reliable results of MGMT assessment in pituitary adenomas.

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Title: MGMT assessment in pituitary adenomas: comparison of different immunohistochemistry fixation chemicals
Authors: Alexander S. G. Micko
Romana Höftberger
Adelheid Wöhrer
Matthias Millesi
Engelbert Knosp
Stefan Wolfsberger
Publication date: 01-06-2018
Publisher: Springer US
Published in: Pituitary / Issue 3/2018
Print ISSN: 1386-341X
Electronic ISSN: 1573-7403
DOI: https://doi.org/10.1007/s11102-018-0862-x

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