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3 H‐thymidine is a defective tool with which to measure rates of DNA synthesis
Author(s) -
Hu Valerie W.,
Black Gavin E.,
TorresDuarte Armida,
Abramson Fred P.
Publication year - 2002
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/fj.02-0142fje
Subject(s) - thymidine , dna synthesis , dna replication , dna , cell cycle , chemistry , cell growth , microbiology and biotechnology , cell , biochemistry , biology , radiochemistry
Metabolic incorporation of 3 H‐thymidine into cellular DNA is a widely used protocol to monitor rates of DNA synthesis and cell proliferation. However, this radiochemical has also been reported to induce cell‐cycle arrest and apoptosis in addition to DNA damage. Using stable isotope‐labeled thymidine, we demonstrate that 3 H‐thymidine induces dose‐dependent inhibition of the rate of DNA synthesis. This inhibition occurred within the first round of replication after addition of the radiolabeled tracer and demonstrates the cytotoxic effects of conventional doses of 3 H‐thymidine (typically ≥ 1 μCi/ml). These results thus show that stable isotope methods are superior to radioisotopes for determining rates of DNA synthesis and cell replication. Because 3 H‐thymidine perturbs the very process it was employed to study, experiments using 3 Hthymidine to monitor DNA synthesis and cell proliferation should be interpreted with caution.