Fluorescent reporter assays provide direct, accurate, quantitative measurements of MGMT status in human cells.
Zachary D NagelAndrew A BeharryPatrizia MazzucatoGaspar J KitangeJann N SarkariaEric T KoolLeona D SamsonPublished in: PloS one (2019)
The DNA repair protein O6-methylguanine DNA methyltransferase (MGMT) strongly influences the effectiveness of cancer treatment with chemotherapeutic alkylating agents, and MGMT status in cancer cells could potentially contribute to tailored therapies for individual patients. However, the promoter methylation and immunohistochemical assays presently used for measuring MGMT in clinical samples are indirect, cumbersome and sometimes do not accurately report MGMT activity. Here we directly compare the accuracy of 6 analytical methods, including two fluorescent reporter assays, against the in vitro MGMT activity assay that is considered the gold standard for measuring MGMT DNA repair capacity. We discuss the relative advantages of each method. Our data indicate that two recently developed fluorescence-based assays measure MGMT activity accurately and efficiently, and could provide a functional dimension to clinical efforts to identify patients who are likely to benefit from alkylating chemotherapy.
Keyphrases
- dna repair
- end stage renal disease
- high throughput
- chronic kidney disease
- ejection fraction
- newly diagnosed
- crispr cas
- peritoneal dialysis
- dna methylation
- systematic review
- gene expression
- high resolution
- transcription factor
- squamous cell carcinoma
- single molecule
- dna damage response
- machine learning
- radiation therapy
- genome wide
- mass spectrometry
- single cell
- binding protein
- big data
- amino acid
- circulating tumor cells
- silver nanoparticles