Cellular uptake and antiproliferative effects of 11-oxo-eicosatetraenoic acid.
Cellular uptake and antiproliferative effects of 11-oxo-eicosatetraenoic acid.
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DOI：
10.1194/jlr.m040741
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发表时间：
2013-11
影响因子：
6.5
通讯作者：
Blair IA
中科院分区：
文献类型：
--
作者：
Snyder NW;Revello SD;Liu X;Zhang S;Blair IA
环加氧酶（考克斯）将花生四烯酸（AA）代谢为羟基二十碳四烯酸（HETE），然后其可以被脱氢酶如15-羟基前列腺素脱氢酶（15-PGDH）氧化为氧代-二十碳四烯酸（埃特）。我们先前已经确定11-氧代-二十碳四烯酸（氧代-ETE）和15-氧代-ETE是考克斯-2/15-PGDH衍生代谢物。稳定同位素稀释（SID）手性液相色谱结合电子捕获大气压化学电离（ECAPCI）单反应监测（SRM）MS已用于定量LoVo细胞和人脐静脉内皮细胞（HUVEC）中11-氧代-ETE和15-氧代-ETE的摄取。细胞内11-氧代-和15-氧代-ETE浓度在1小时内达到最高水平，并迅速下降，LoVo细胞和HUVEC之间的摄取存在显著的定量差异。11-oxo-ETE的最大细胞内浓度在LoVo细胞中为0.02 ng/4 × 105个细胞，在HUVEC中为0.58 ng/4 × 105个细胞。相反，LoVo细胞中15-oxo-ETE的最大水平为0.21 ng/4 × 105，HUVEC中为0.01 ng/4 × 105。11-氧代-和15-氧代-ETE的甲酯使相应游离氧代-ETE的细胞内浓度增加3至8倍。11-oxo-ETE、15-oxo-ETE及其甲酯在2-10 μM浓度下抑制HUVEC和LoVo细胞的增殖，其中11-oxo-ETE甲酯是最有效的抑制剂。与多药耐药转运蛋白（MRP）1和4抑制剂丙磺舒共同处理，11-oxo-ETE甲酯在LoVo细胞中的抗增殖作用增强，11-oxo-ETE的细胞内浓度从0.05 ng/4 × 105细胞增加到0.18 ng/4 × 105细胞。因此，本研究已经确定考克斯-2/15-PGDH衍生的类二十烷酸11-氧代-和15-氧代-ETE进入靶细胞，它们抑制细胞增殖，并且它们的抑制作用由MRP输出物调节。
Cyclooxygenases (COX) metabolize arachidonic acid (AA) to hydroxyeicosatetraenoic acids (HETE), which can then be oxidized by dehydrogenases, such as 15-hydroxyprostaglandin dehydrogenase (15-PGDH), to oxo-eicosatetraenoic acids (ETE). We have previously established that 11-oxo-eicosatetraenoic acid (oxo-ETE) and 15-oxo-ETE are COX-2/15-PGDH-derived metabolites. Stable isotope dilution (SID) chiral liquid chromatography coupled with electron capture atmospheric pressure chemical ionization (ECAPCI) single reaction monitoring (SRM) MS has been used to quantify uptake of 11-oxo-ETE and 15-oxo-ETE in both LoVo cells and human umbilical vein endothelial cells (HUVEC). Intracellular 11-oxo- and 15-oxo-ETE concentrations reached maximum levels within 1 h and declined rapidly, with significant quantitative differences in uptake between the LoVo cells and the HUVECs. Maximal intracellular concentrations of 11-oxo-ETE were 0.02 ng/4 × 105 cells in the LoVo cells and 0.58 ng/4 × 105 cells in the HUVECs. Conversely, maximal levels of 15-oxo-ETE were 0.21 ng/4 × 105 in the LoVo cells and 0.01 ng/4 × 105 in the HUVECs. The methyl esters of both 11-oxo- and 15-oxo-ETE increased the intracellular concentrations of the corresponding free oxo-ETEs by 3- to 8-fold. 11-oxo-ETE, 15-oxo-ETE, and their methyl esters inhibited proliferation in both HUVECs and LoVo cells at concentrations of 2–10 μM, with 11-oxo-ETE methyl ester being the most potent inhibitor. Cotreatment with probenecid, an inhibitor of multiple drug resistance transporters (MRP)1 and 4, increased the antiproliferative effect of 11-oxo-ETE methyl ester in LoVo cells and increased the intracellular concentration of 11-oxo-ETE from 0.05 ng/4 × 105 cells to 0.18 ng/4 × 105 cells. Therefore, this study has established that the COX-2/15-PGDH-derived eicosanoids 11-oxo- and 15-oxo-ETE enter target cells, that they inhibit cellular proliferation, and that their inhibitory effects are modulated by MRP exporters.