Valentin I, Philippe M, Lhuguenot J, Chagnon M
ENSBANA, UMR (O938) Toxicologie Alimentaire, 1 Esplanade Erasme, 21000, Dijon, France.
Toxicology. 2001 Feb 14;158(3):127-39. doi: 10.1016/s0300-483x(00)00372-3.
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.
本研究描述了一种基于HepG2细胞中RNA合成抑制程度来测量细胞毒性的灵敏微量测定法。RNA合成通过放射性标记尿苷的动力学摄取来测量。在广泛的浓度范围内测试了大量化合物。确定每种化合物诱导HepG2尿苷摄取率50%抑制所需的浓度（IC50），并用于对其效力进行排名。将这些IC50与用中性红试验测得的IC50进行比较。在中性红试验中，2-乙酰氨基芴、苯并[a]芘和甲基亚硝基脲无细胞毒性。尿苷摄取总是在低于中性红试验所需的浓度下受到抑制，这表明尿苷摄取试验比中性红包涵体试验更灵敏地指示毒性作用。尿苷摄取试验提供了一种快速定量的方法来评估人类细胞系中的毒性。描述了该方法在瓶装矿泉水中的应用。由于其高灵敏度和可重复性，该方法为筛选大量样品提供了合适的工具，并且将成为评估食品安全和控制包装材料回收过程的有用测试。
