A Fatal Combination: A Thymidylate Synthase Inhibitor with DNA Damaging Activity

• Published in: PloS one Vol. 10; no. 2; p. e0117459
• Main Authors: Ligasová, Anna, Strunin, Dmytro, Friedecký, David, Adam, Tomáš, Koberna, Karel
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 11.02.2015; Public Library of Science (PLoS)
• Subjects: Abnormalities; Analysis; Biochemistry; Cancer; Cell cycle; Cell death; Cell division; Cell growth; Cell Line; Cell lines; Cell Proliferation - drug effects; Crosslinking; Cytotoxicity; Cytotoxins - metabolism; Cytotoxins - toxicity; Damage; Deoxyribonucleic acid; Deoxyuridine - analogs & derivatives; Deoxyuridine - metabolism; Deoxyuridine - toxicity; Dihydrofolate reductase; DNA; DNA - biosynthesis; DNA - genetics; DNA - metabolism; DNA Damage; DNA Replication - drug effects; Dose-Response Relationship, Drug; Enzyme Inhibitors - metabolism; Enzyme Inhibitors - toxicity; Herpes viruses; Humans; Inhibition; Intracellular Space - drug effects; Intracellular Space - metabolism; Kinases; Mass spectrometry; Medicine; Metabolism; Metabolites; Physiological aspects; S phase; S Phase - drug effects; Scientific imaging; Synthesis; Tetrahydrofolates - biosynthesis; Thymidine - metabolism; Thymidine - pharmacology; Thymidine monophosphate; Thymidine Monophosphate - metabolism; Thymidylate synthase; Thymidylate Synthase - antagonists & inhibitors; Toxicity; Hungary; Czech Republic
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0117459

2'-Deoxy-5-ethynyluridine (EdU) has been previously shown to be a cell poison whose toxicity depends on the particular cell line. The reason is not known. Our data indicates that different efficiency of EdU incorporation plays an important role. The EdU-mediated toxicity was elevated by the inhibition of 2'-deoxythymidine 5'-monophosphate synthesis. EdU incorporation resulted in abnormalities of the cell cycle including the slowdown of the S phase and a decrease in DNA synthesis. The slowdown but not the cessation of the first cell division after EdU administration was observed in all of the tested cell lines. In HeLa cells, a 10 μM EdU concentration led to the cell death in the 100% of cells probably due to the activation of an intra S phase checkpoint in the subsequent S phase. Our data also indicates that this EdU concentration induces interstrand DNA crosslinks in HeLa cells. We suppose that these crosslinks are the primary DNA damage resulting in cell death. According to our results, the EdU-mediated toxicity is further increased by the inhibition of thymidylate synthase by EdU itself at its higher concentrations.