人癌细胞中3-脱氮尿苷-5'-三磷酸的高压液相色谱分析
High-pressure liquid chromatographic analysis of 3-deazauridine-5'-triphosphate in human cancer cells.
作者信息
Plunkett W, Benvenuto J A, Stewart D J, Loo T L
出版信息
Cancer Treat Rep. 1979 Mar;63(3):415-20.
A rapid high-pressure liquid chromatographic assay for the detection and quantitation of 3-deazauridine-5'-triphosphate (deazaUTP), the active metabolite of the anticancer drug 3-deazauridine (deazaUrd), in cell extracts is described. This procedure permits the simultaneous detection and quantitation of CTP, the cellular concentration of which is affected by treatment with deazaUrd. Human lymphoblastoid cells (line CCRF-CEM) treated with 100 nmols of deazaUrd/ml in culture accumulate deazaUTP to greater than 25 nmols/10(7) cells after 4 hours. The concentration of CTP in these cells decreased exponentially with a half-life of 1.1 hours. After these cells had been incubated with 100 nmols of deazaUrd/ml and were resuspended in drug-free medium, the intracellular deazaUTP concentration decreased exponentially with a half-life of 3.4 hours. This assay has been applied to clinical studies of deazaUrd; high cellular deazaUTP concentrations have been detected in brain tumor tissue after deazaUrd infusion.
本文描述了一种快速高压液相色谱分析法，用于检测和定量细胞提取物中的3 - 去氮尿苷 - 5'- 三磷酸（deazaUTP），它是抗癌药物3 - 去氮尿苷（deazaUrd）的活性代谢产物。该方法可同时检测和定量CTP，其细胞浓度会受到deazaUrd处理的影响。在培养中用100 nmol/ml的deazaUrd处理的人淋巴母细胞系（CCRF - CEM），4小时后积累的deazaUTP大于25 nmol/10⁷个细胞。这些细胞中CTP的浓度呈指数下降，半衰期为1.1小时。在用100 nmol/ml的deazaUrd孵育这些细胞并将其重悬于无药物培养基后，细胞内deazaUTP浓度呈指数下降，半衰期为3.4小时。该分析方法已应用于deazaUrd的临床研究；在输注deazaUrd后，在脑肿瘤组织中检测到高细胞deazaUTP浓度。
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