Uridine kinase activities and pyrimidine nucleoside phosphorylation in fluoropyrimidine-sensitive and -resistant cell lines of the Novikoff hepatoma.
Uridine kinase, the rate-limiting enzyme in the activation (phosphorylation) of uridine and the corresponding chemotherapeutic analogues, is present as two isoenzymes localized exclusively in the cytosol of rapidly growing neoplasms, including the S-37 sarcoma, EL-4 leukaemia, HeLa cells (a human carcinoma) and the Novikoff hepatoma. The activities of the isolated isoenzymes are markedly decreased when the concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations approximating to those found in vivo. Further, comparisons of the Km values of isolated uridine kinases with those for cellular uptake of pyrimidine nucleosides and their rate of intracellular phosphorylation suggest that nucleoside-transport systems play a rate-limiting role in nucleoside analogue activation and consequently that it is impossible to estimate the Km of uridine kinase in the intact cell. During the development of tumour-cell resistance to 5-fluorouracil or 5-fluorouridine in vivo there was an early differential increase in the activity of a low-affinity (high-Km) uridine kinase isoenzyme, as measured in cell extracts, and a 7-fold increase in the Km values for the uptake of both uridine and 5-fluorouridine into the intact resistant cells.
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000378:metabolism*; D002460:Cell Line; D004351:Drug Resistance; D005260:Female; D005472:Fluorouracil / Q000494:pharmacology*; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D008113:Liver Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D008274:Magnesium / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects; D009374:Neoplasms, Experimental / Q000378:metabolism; D010710:Phosphates; D010770:Phosphotransferases / Q000378:metabolism*; D011741:Pyrimidine Nucleosides / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology; D014541:Uridine Kinase / Q000378:metabolism*
D007527:Isoenzymes; D010710:Phosphates; D011741:Pyrimidine Nucleosides; D010770:Phosphotransferases; D014541:Uridine Kinase; D008274:Magnesium; D005472:Fluorouracil