Untargeted metabolomics profiles delineate metabolic alterations in mouse plasma during lung carcinoma development using UPLC-QTOF/MS in MSE mode

In this work, an untargeted metabolomic method based on ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) in MSE (E represents collision energy) mode was exploited to determine the dynamic metabolic alterations in the plasma of male C57BL/6 mice during the onset and development of lung carcinoma. Plasma samples were collected from control and model mice (male C57BL/6 mice experimentally inoculated with the Lewis lung carcinoma cells) at 7 and 14 days post-inoculation (DPI). As a result, 15 dysregulated metabolites, including cholesterol sulphate, tiglylcarnitine, 1-palmitoylglycerophosphoinositol, 2-stearoylglycerophosphoinositol, stearoylcarnitine, PC(20:2(11Z,14Z)/16:0), PC(22:4(7Z,10Z,13Z,16Z)/14:0), PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:0), PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:0), 12,20-Dioxo-leukotriene B4, sphingosine 1-phosphate(d19:1-P), sphingomyelin(d18:0/16:1(9Z)), lysoPC(16:0), lysoPC(18:0) and lysoPC(20:4(5Z,8Z,11Z,14Z)), were identified in the plasma of model mice with xenografts at both 7 and 14 DPI. All the altered metabolites associated with the onset and development of lung carcinoma were involved in the metabolism of glycerophospholipid, fatty acid, sphingolipid and arachidonic acid. The feasible utility of these endogenous biomarkers as potential diagnostic indicators was validated through receiver operating characteristic curve analysis. Collectively, these findings provide a systematic view of metabolic changes linked to the onset and development of lung carcinoma.

本研究采用基于超高效液相色谱-四极杆-飞行时间质谱（ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry，UPLC-QTOF/MS）的非靶向代谢组学方法，在MSE模式（其中E代表碰撞能量）下，检测雄性C57BL/6小鼠肺癌发生发展过程中血浆的动态代谢变化。本研究在接种后第7天和第14天（DPI），分别从对照组小鼠与模型组小鼠——即经实验接种Lewis肺癌细胞的雄性C57BL/6小鼠——采集血浆样本。最终，在接种后第7天和第14天的异种移植模型小鼠血浆中，共鉴定出15个差异代谢物，包括硫酸胆固醇、巴豆酰肉碱、1-棕榈酰甘油磷酸肌醇、2-硬脂酰甘油磷酸肌醇、硬脂酰肉碱、磷脂酰胆碱（Phosphatidylcholine，PC）(20:2(11Z,14Z)/16:0)、PC(22:4(7Z,10Z,13Z,16Z)/14:0)、PC(22:5(7Z,10Z,13Z,16Z,19Z)/14:0)、PC(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:0)、12,20-二氧代白三烯B4、1-磷酸鞘氨醇(d19:1-P)、鞘磷脂(d18:0/16:1(9Z))、溶血磷脂酰胆碱（lysophosphatidylcholine，lysoPC）(16:0)、lysoPC(18:0)及lysoPC(20:4(5Z,8Z,11Z,14Z))。所有与肺癌发生发展相关的差异代谢物均参与了甘油磷脂、脂肪酸、鞘磷脂及花生四烯酸的代谢过程。通过受试者工作特征（receiver operating characteristic，ROC）曲线分析，验证了这些内源性生物标志物作为潜在诊断指标的应用可行性。综上，本研究结果系统阐明了与肺癌发生发展相关的代谢变化规律。