Core--Eicosanoid Analysis

描述：（申请人描述）类二十烷酸分析核心的具体目标是为本计划项目资助的研究人员进行复杂生物流体和组织中的类二十烷酸分析。本计划计划资助的四个项目涉及研究环氧化酶和类二十烷酸在结直肠肿瘤中的作用，并将需要量化前列腺素（pg）。在细胞培养和动物及人体组织中，将采用毛细管气相色谱(GC)/电子捕获负离子化学电离（NICI）质谱（MS）的方法，使用由核心领导者开发并目前正在使用的稳定同位素稀释技术，对类二十烷进行定量。参与本项目资助的研究人员使用这些检测方法已经进行了一些基本观察，这些观察有助于描述类二十烷在胃肠道恶性肿瘤相关病理生理过程中的作用。此外，Core还将开发一种GC/NICI质谱法来测定化合物12-羟基十七碳三烯酸（12- hht），该化合物来源于环加氧酶生成的双环烯过氧化物中间体PGH2。该试验将用于项目4 (Dr. Marnett)，以进一步探索12-HHT在环加氧酶过表达情况下的作用。因此，二十烷类核心的总体目标是为二十烷类的定量分析提供技术支持和高灵敏度方法的实施。

DESCRIPTION: (Applicant's Description) The specific goals of the Eicosanoid Analysis Core are to carry out the analyses of eicosanoids in complex biological fluids and tissues for investigators in this Program Project Grant. The four projects proposed in this Program Project Grant involve research examining the role of the cyclooxygenase and eicosanoids in colorectal neoplasia and will require quantification of prostaglandins (PGs). Eicosanoids will be quantified in cellular incubations and animal and human tissues by methodology employing capillary gas chromatography (GC)/electron capture negative ion chemical ionization (NICI) mass spectrometry (MS) using stable isotope dilution techniques that were developed by the Core Leader and which are currently in use. Use of these assays by investigators involved in this Program Project Grant has already allowed a number of fundamental observations to be made that have helped to delineate the role of eicosanoids in pathophysiological processes related to gastrointestinal malignancy. In addition, the Core will also develop a GC/NICI MS assay to measure the compound 12- hydroxy-heptadecatrienoic acid (12-HHT) derived from the cyclooxygenase-generated bicycloendoperoxide intermediate PGH2. This assay will be utilized in Project 4 (Dr. Marnett) to further explore the role of 12-HHT in situations of cyclooxygenase overexpression. The overall objectives of the Eicosanoid Core, therefore, are to provide technical support and implementation of highly sensitive methodology for the quantitative analysis of eicosanoids.