Uridine uptake inhibition as a cytotoxicity test for a human hepatoma cell line (HepG2 cells): comparison with the neutral red assay.
This study describes a sensitive microassay for measuring cytotoxicity based on the degree of inhibition of RNA synthesis in HepG2 cells. RNA synthesis is measured by the kinetic uptake of radiolabeled uridine. A large number of compounds were tested in a wide range of concentrations. The concentration required to induce 50% inhibition of HepG2 uridine uptake rates (IC(50)) was determined for each compound and used to rank its potency. These IC(50)s were compared with IC(50)s measured with the neutral red assay. 2-acetylaminofluorene, benzo[a]pyrene and methylnitrosourea were not cytotoxic in the neutral red assay. Uridine uptake was always inhibited at lower concentrations than those required in the neutral red assay, suggesting that the uridine uptake assay is a more sensitive indicator of toxic action than the neutral red inclusion. Uridine uptake assay provides a rapid and quantitative method for assessing toxicity in a human cell line. Application of this method to bottled spring waters are described. Due to its high sensitivity and reproducibility, this method provides a suitable tool for screening a great number of samples and will be a helpful test for evaluating food safety and controlling the recycling process of wrapping materials.
ENSBANA, UMR (O938) Toxicologie Alimentaire, 1 Esplanade Erasme, 21000, Dijon, France.
I
I
Valentin
M
M
Philippe
J
J
Lhuguenot
M
M
Chagnon
D015073:2-Acetylaminofluorene / Q000633:toxicity; D001580:Benzopyrenes / Q000633:toxicity; D002273:Carcinogens / Q000633:toxicity*; D006528:Carcinoma, Hepatocellular; D004396:Coloring Agents / Q000737:chemistry; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008770:Methylnitrosourea / Q000633:toxicity; D009499:Neutral Red / Q000737:chemistry; D012313:RNA / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry; D012044:Regression Analysis; D015203:Reproducibility of Results; D018675:Toxicity Tests / Q000379:methods*; D014407:Tumor Cells, Cultured; D014529:Uridine / Q000493:pharmacokinetics*; D014874:Water Pollutants, Chemical / Q000032:analysis; D015262:Xenobiotics / Q000633:toxicity*
D001580:Benzopyrenes; D002273:Carcinogens; D004396:Coloring Agents; D014874:Water Pollutants, Chemical; D015262:Xenobiotics; D009499:Neutral Red; D012313:RNA; D008770:Methylnitrosourea; D015073:2-Acetylaminofluorene; D014529:Uridine