Novel dynamic flow cytoenzymological determination of intracellular esterase inhibition by BCNU and related isocyanates.
Novel dynamic flow cytoenzymological determination of intracellular esterase inhibition by BCNU and related isocyanates.
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BCNU 和相关异氰酸酯抑制细胞内酯酶的新型动态流式细胞酶学测定。
DOI：
10.1016/0006-2952(87)90027-x
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发表时间：
1987
影响因子：
5.8
通讯作者：
J. Watson
中科院分区：
文献类型：
--
作者：
C. Dive;P. Workman;J. Watson
我们提出了一种新的动态流动细胞酶学方法，证明了抗癌药物氯乙基亚硝酸BCNU能有效地抑制活的完整小鼠和人肿瘤细胞的酯酶对二乙酸荧光素的细胞内水解酶的作用。BCNU偏苯二甲酸氯乙基异氰酸酯和相关的复合异氰酸酯也是有效的缓蚀剂，其I50值在4.2×10−5~2.0×10−4M之间。常规荧光分光光度法对完整细胞和声酸盐的定量结果基本相似，并证明纯化的猪肝羧基酯酶(EC 3.1.1.1)具有抑制作用。烷基化试剂甲基甲烷磺酸盐、马法兰和氮芥末几乎没有或几乎没有抑制活性。这一结果与异氰酸酯对酯酶分子的氨基甲酰化反应一致，这可能为异质样品的离散亚群中细胞内氨基甲酰化反应的流式细胞术测定提供了基础。
We present a novel dynamic flow cytoenzymological demonstration of the potent inhibition by the antitumour chloroethylnitrosourea BCNU of the intracellular hydrolysis of fluorescein diacetate by esterases of viable, intact murine and human tumour cellsin vitro. The BCNU metabonate chloroethyl isocyanate and the related compoundn-butyl isocyanate were also potent inhibitors.I50values were in the range 4.2 × 10−5–2.0× 10−4M. Generally similar quantitative results were obtained for intact cells and sonicates by conventional spectrofluorimetry, and inhibition of purified porcine liver carboxyl esterase (EC 3.1.1.1) was demonstrated. Little or no inhibitory activity was seen with the alkylating agents methyl methane-sulphonate, melphalan and nitrogen mustard. The results are consistent with carbamoylation of the esterase molecules by isocyanates, and this may provide a basis for the flow cytometric determination of intracellular carbamoylation in discrete subpopulations of heterogeneous samples.