Wickramasinghe S N, Fida S
Dept. of Haematology, St. Mary's Hospital Medical School, Imperial College of Science, Technology & Medicine, London, U.K.
Eur J Haematol. 1993 Mar;50(3):127-32. doi: 10.1111/j.1600-0609.1993.tb00080.x.
HL60 cells were cultured for 10 days under various experimental conditions. They were then incubated with 1 mumol/l [5-3H] uridine for 2 hours and their DNA extracted. The DNA was hydrolysed to deoxyribonucleosides with phosphodiesterase and alkaline phosphatase and the hydrolysate subjected to Aminex A6 chromatography. The elution profiles showed that, when compared with control cells, DNA from cells grown in medium deficient in folate, B12 or both folate and B12 contained increased amounts of deoxyuridine (dU) and increased radioactivity in the dU peak. The data demonstrate that misincorporation of uracil into DNA occurs in a myeloid cell line cultured in growth medium deficient in folate, B12 or both folate and B12.
HL60细胞在各种实验条件下培养10天。然后将它们与1微摩尔/升的[5-³H]尿苷孵育2小时,并提取其DNA。用磷酸二酯酶和碱性磷酸酶将DNA水解为脱氧核糖核苷,并将水解产物进行Aminex A6色谱分析。洗脱图谱显示,与对照细胞相比,在缺乏叶酸、维生素B12或叶酸和维生素B12两者的培养基中生长的细胞的DNA含有增加量的脱氧尿苷(dU),并且dU峰中的放射性增加。数据表明,在缺乏叶酸、维生素B12或叶酸和维生素B12两者的生长培养基中培养的髓系细胞系中,尿嘧啶错误掺入DNA中。