Metabolomics of adherent mammalian cells by capillary electrophoresis-mass spectrometry: HT-29 cells as case study

摘要: Abstract In this work, the optimization of an effective protocol for cell metabolomics is described with special emphasis in sample preparation and subsequent analysis intracellular metabolites from adherent mammalian cells by capillary electrophoresis-mass spectrometry. As case study, colon cancer HT-29 cells, a human model to investigate cancer, are employed. The feasibility whole method demonstrated via fast sensitive profiling electrophoresis-time-of-flight mass spectrometry (CE-TOF MS). suitability methodology further corroborated through examination metabolic changes polyamines pathway produced difluoromethylornithine (DFMO), known potent ornithine decarboxylase inhibitor. selection optimum extraction conditions allowed higher volume injection that led increase CE-TOF MS sensitivity. Following non-targeted approach, 10 (namely, putrescine, ornithine, gamma-aminobutyric acid (GABA), oxidized reduced glutathione, 5′-deoxy-5′-(methylthio)adenosine, N-acetylputrescine, cysteinyl-glycine, spermidine unknown compound) were found be significantly altered DFMO ( p