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Development of a mass spectrometry assay to measure ribonucleotide reductase activity in human cancer cells
Author(s) -
Tong Ai Phuong S.,
Rice Kevin P
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.560.8
Subject(s) - ribonucleotide reductase , thioredoxin reductase , cancer cell , chemistry , reductase , biochemistry , cancer , prodrug , deoxyribonucleotides , chronic myelogenous leukemia , dna replication , enzyme , cancer research , biology , leukemia , dna , thioredoxin , immunology , genetics , gene , nucleotide , protein subunit
Ribonucleotide reductase (RNR) catalyzes the reduction of ribonucleotides into deoxyribonucleotides, which are essential for DNA replication and cell division. RNR is especially abundant in proliferating cancer cells and is an increasingly popular target in anticancer research. We had previously reported that the experimental anticancer prodrug Laromustine inhibits the enzyme thioredoxin reductase (TrxR), which is involved in proper RNR activity. Laromustine has been used in clinical trials to treat patients with acute myelogenous leukemia and glioblastoma multiforme. In this study, we use a particular sample preparation method and HPLC coupled to high‐resolution time of flight mass spectrometry to quantify deoxyribonucledotide diphosphate pools in human cells. These experiments will allow us to assess changes in RNR activity from cultured human cancer cells in response to treatment with Laromustine. We hope to further elucidate the mechanism by which Laromustine kills cancer cells, possibly influencing the clinical applications of Laromustine or similar agents.