The Study Of Amino Acid Metabolic Footprinting Analysis Of Lung Cancer Cells By Liquid Chromatography-tandem Mass Spectrometry

Metabolomics can observe the metabolites of changes in biological systems perturbed by environmental factors and genetic factors,which play an important role in systems biology.Cell metabolomics include metabolic fingerprinting and metabolic footprinting analysis,wherein metabolic footprinting analysis is used to characterize metabolic pathways in cultured cell lines by analyzing nutrient consumption and metabolite secretion of cells,and find out the pivotal biomarkers for kinetic and non-invasive monitoring of cell metabolism.To explore the distinctive profiles of lung cancer cell lines and define the metabolic phenotypes of the cell lines,our research has developed a footprinting analysis based on LC-QTOF using non-targeted metabolomics for different metabolites between lung cancer and non-cancer cell lines.Furthermore,chemometrics and bioinformatics were also applied.Then,characteristic metabolites between lung cancer and non-cancer cell lines in consecutive days were found.Moreover,based on preliminary research of our group and combined with the studies of amino acid metabolism of tumor,we have developed a new method using stable isotope-labeled(SIL)analogs as the surrogate of amino acids coupled with a HILIC-MS/MS method for the determination of targeted amino acids in cell cultured media.The surrogate analyte approach eliminates the inteRFerence from the endogenous substances in the authentic matrix,making it possible to prepare calibration standards in the real matrix.Besides,this method avoided the usage of derivatization and ion-pairing reagents by employing HILIC.The method has been successfully used to monitor the dynamic changes of amino acid release or uptake in lung cancer and non-cancer cell lines,and to investigate amino acid profiles of lung cancer cell lines.In summary,this study aims to develop metabolic footprinting analysis based on LC-MS in lung cancer and non-cancer cell lines,obtain different amino acid metabolic profiles between them and provide valuable clues for discovering the biomarker and developing methodological techniques for amino acid metabolic studies of cancer cells.