Use of electroporation to study the cytotoxic effects of fluorodeoxyuridylate in intact cells.
The introduction of 2'-deoxyuridine 5'-monophosphate and its analog, 5-fluoro-2'-deoxyuridine 5'-monophosphate, into intact CCRF-CEM and NIH3T3 cells was achieved by electroporation. Following electroporation, cells were shown to be fully functional as monitored by the incorporation of deoxyuridylate, after conversion to thymidylate, into DNA. Pretreatment of cells with fluorodeoxyuridine completely abolished this effect. In contrast, introduction of the fluoro analog into cells by electroporation markedly inhibited both DNA synthesis and cell growth in a time-dependent manner. Thus, electroporation offers a powerful tool to permeabilize cells to a variety of cellular metabolites and antimetabolites.
D002460:Cell Line; D002463:Cell Membrane Permeability; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000494:pharmacology*; D003857:Deoxyuridine / Q000378:metabolism; D005468:Fluorodeoxyuridylate / Q000378:metabolism / Q000494:pharmacology*; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors
D003856:Deoxyuracil Nucleotides; D005468:Fluorodeoxyuridylate; D004247:DNA; D013940:Thymidylate Synthase; D003857:Deoxyuridine