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垂体腺瘤中 MGMT 的评估：不同免疫组织化学固定化学试剂的比较。

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

机构信息

Department of Neurosurgery, Medical University Vienna, Waehringer Guertel 18-20, 1097, Vienna, Austria.

Institute of Neurology, Medical University of Vienna, Vienna, Austria.

出版信息

Pituitary. 2018 Jun;21(3):266-273. doi: 10.1007/s11102-018-0862-x.

DOI:10.1007/s11102-018-0862-x
PMID:29344904
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5942339/
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.

摘要

目的

尽管 O6-甲基鸟嘌呤-DNA 甲基转移酶（MGMT）已被确立为替莫唑胺反应的标志物，但评估垂体腺瘤中 MGMT 表达的最可靠方法仍存在争议。目前，甲醛固定组织样本的免疫组织化学（IHC）评估在半定量描述中应用最广泛。由于甲醛不能完全保存核酸，因此提出了 RCL2（一种无甲醛的酒精固定剂）作为更可靠的替代物，在细胞稳定性方面具有优势。此外，由于当前的 IHC 方法是半定量和依赖观察者的，因此出现了一种客观的评估 MGMT 启动子甲基化状态的工具——焦磷酸测序，作为一种可靠和准确的替代方法。本研究旨在验证当前用于评估垂体腺瘤中 MGMT 蛋白表达的 IHC 方法。

方法

对 8 例具有阳性 IHC MGMT 表达（>50%）的大腺瘤组织样本进行了研究：首先，我们比较了甲醛与 RCL2 固定后垂体腺瘤切除后 MGMT 蛋白表达的时间依赖性稳定性。然后，我们使用焦磷酸测序比较了阳性 IHC MGMT 表达与甲基化启动子状态。

结果

在腺瘤切除后的头 12 小时内，使用甲醛固定的组织样本中 MGMT 阳性的比例明显高于 RCL2 固定的组织样本（分别为 96%和 81%，p=0.025）。

结论

我们的数据证实，目前使用甲醛组织固定和 IHC 的方法在评估垂体腺瘤中的 MGMT 方面显示出稳定可靠的结果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/66be148ff8d9/11102_2018_862_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/01acd0221563/11102_2018_862_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/d11113a1806f/11102_2018_862_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/c0fd1fa06537/11102_2018_862_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/66be148ff8d9/11102_2018_862_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/01acd0221563/11102_2018_862_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/d11113a1806f/11102_2018_862_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/c0fd1fa06537/11102_2018_862_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a9c4/5942339/66be148ff8d9/11102_2018_862_Fig4_HTML.jpg

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