Artikel
RNAscope: a new method to detect MGMT status?
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Veröffentlicht: | 25. August 2015 |
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Gliederung
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MGMT promoter methylation is a strong prognostic biomarker in glioblastoma patients treated with temozolomide. Methylation-specific PCR and pyrosequencing are the most commonly used DNA-based diagnostic methods for promoter methylation. Both are time- and cost-intensive methods harboring limitations, e.g. due to amount of tumor tissue and bisulfite treatment.
In situ hybridization is an extremely powerful research tool for gene expression on DNA level and widely used in routine diagnostics, e.g. HER2-neu amplification status. Especially chromogenic in situ hybridization is a fast and cost-effective method, which could be easily evaluated on bright field microscopy.
RNAscope is a new and extremely sensitive, in situ hybridization based method to visualize RNA molecules, which is compatible with fresh frozen and formalin fixed paraffine embedded tissue and does not require RNA-free environment.
We assume that CpG-Island methylation of MGMT promoter resulting in transcriptional silencing, is mirrowed by lack of MGMT RNA expression, whereas unmethylated tumor tissue shows strong RNA expression visualized by chromogenic in situ hybridization.
Our aim is to investigate whether this bright field based in situ hybridization method might be an achievement to assess MGMT status.