Fluorescent reporter assays provide direct, accurate, quantitative measurements of MGMT status in human cells

Zachary D. Nagel, Andrew A. Beharry, Patrizia Mazzucato, Gaspar J. Kitange, Jann N Sarkaria, Eric T. Kool, Leona D. Samson

Research output: Contribution to journalArticle

Abstract

The DNA repair protein O 6 -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.

Original languageEnglish (US)
Article numbere0208341
JournalPloS one
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2019

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Nagel, Z. D., Beharry, A. A., Mazzucato, P., Kitange, G. J., Sarkaria, J. N., Kool, E. T., & Samson, L. D. (2019). Fluorescent reporter assays provide direct, accurate, quantitative measurements of MGMT status in human cells. PloS one, 14(2), [e0208341]. https://doi.org/10.1371/journal.pone.0208341