Modulation of cellular S1P levels with a novel, potent and specific inhibitor of sphingosine kinase-1
Modulation of cellular S1P levels with a novel, potent and specific inhibitor of sphingosine kinase-1
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DOI：
10.1042/bj20111929
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发表时间：
2012-05-15
影响因子：
4.1
通讯作者：
Nagiec, Marek M.
中科院分区：
文献类型：
--
作者：
Schnute, Mark E.;McReynolds, Matthew D.;Nagiec, Marek M.
SphK（鞘氨醇激酶）是生物活性脂质和GPCR（G蛋白偶联受体）激动剂S1 P（鞘氨醇1-磷酸）的主要来源。S1 P促进细胞生长、存活和迁移，是淋巴细胞运输的关键调节因子。已经提出抑制S1 P信号传导作为治疗炎性疾病和癌症的策略。在本文中，我们描述了PF-543，一种新的细胞渗透性SphK 1抑制剂的发现和表征。PF-543抑制SphK 1的Ki为3.6 nM，具有鞘氨醇竞争性，对SphK 1的选择性是SphK 2亚型的100倍以上。在1483头颈癌细胞中，其特征在于SphK 1表达水平高和S1 P产生率异常高，PF-543使内源性S1 P水平降低10倍，鞘氨醇水平成比例增加。与过去的报告表明，许多癌细胞系的生长是SphK 1依赖性的，SphK 1的特异性抑制对1483细胞的增殖和存活没有影响，尽管细胞S1 P/鞘氨醇比例发生了显着变化。PF-543是全血中S1 P形成的有效抑制剂，表明鞘氨醇激酶的SphK 1亚型是人血液中S1 P的主要来源。PF-543是迄今为止描述的最有效的SphK 1抑制剂，它将有助于剖析SphK 1驱动的S1 P信号传导的特定作用。
SphK (sphingosine kinase) is the major source of the bioactive lipid and GPCR (G-protein-coupled receptor) agonist S1P (sphingosine 1-phosphate). S1P promotes cell growth, survival and migration, and is a key regulator of lymphocyte trafficking. Inhibition of S1P signalling has been proposed as a strategy for treatment of inflammatory diseases and cancer. In the present paper we describe the discovery and characterization of PF-543, a novel cell-permeant inhibitor of SphK1. PF-543 inhibits SphK1 with a K-i of 3.6 nM, is sphingosine-competitive and is more than 100-fold selective for SphK1 over the SphK2 isoform. In 1483 head and neck carcinoma cells, which are characterized by high levels of SphK1 expression and an unusually high rate of S1P production, PF-543 decreased the level of endogenous S1P 10-fold with a proportional increase in the level of sphingosine. In contrast with past reports that show that the growth of many cancer cell lines is SphK1-dependent, specific inhibition of SphK1 had no effect on the proliferation and survival of 1483 cells, despite a dramatic change in the cellular S1P/sphingosine ratio. PF-543 was effective as a potent inhibitor of S1P formation in whole blood, indicating that the SphK1 isoform of sphingosine kinase is the major source of S1P in human blood. PF-543 is the most potent inhibitor of SphK1 described to date and it will be useful for dissecting specific roles of SphK1-driven S1P signalling.