Deep mining and quantification of oxidized cholesteryl esters discovers potential biomarkers involved in breast cancer by liquid chromatography-mass spectrometry

Abstract

Oxidized cholesteryl ester (OxCE) is produced by the oxidation of cholesteryl ester (CE) in the cores of lipoproteins. OxCE production and oxidative stress have been largely associated with breast cancer. Herein, we developed a novel reverse-phase liquid chromatography coupling quadrupole time-of-flight mass spectrometry (RPLC‒Q-TOF‒MS) method based on the iterative acquisition mode and used the MS/MS mode for deep mining and simultaneous quantification of cholesterol (Chol), CEs and OxCEs in human serum. A mathematical model was used to globally profile 57 molecular species of both CEs and OxCEs in the serum of both healthy volunteers and patients with breast cancer, and the qualitative results were verified based on the retention regularity. An abnormal elevation of OxCEs was found in serum samples of breast cancer patients, where OxCEs were produced by the oxidation of the fatty acyl chain of CE (20:4), such as CE (20:1)+3O, CE (20:2)+2O and CE (20:3)+O, which could be regarded as biomarkers. This comprehensive method for the global profiling of Chol, OxCEs and CEs sheds light on the role OxCEs and CEs play in breast cancer and has enabled the discovery of breast cancer biomarkers.