AGS vs HGC-27 gastric metabolome-lipidome
Description
Abstract Background Gastric cancer cell lines differ in metabolite and lipid composition, and resolving those differences requires polar and lipid measurements made in parallel on the same material. We compared intracellular and conditioned-medium profiles of AGS and HGC-27 cells. Methods and Results Targeted liquid chromatography–tandem mass spectrometry was applied to four biological replicates per cell line, grown in separate Petri dishes in parallel in cell-line-specific basal media. The retained intracellular panels comprised 31 lipid species and 188 polar features. Principal component analysis separated the two lines completely in all analytical layers. Seven lipid species and 162 polar features reached adjusted p values below 0.05; four and 95, respectively, also met predefined fold-change and standardised-effect thresholds. AGS held higher phosphoethanolamine, lower-glycolytic intermediates, purines, diacylglycerols and cholesteryl esters. HGC-27 held higher mevalonate, oxidised glutathione, 2-hydroxyglutarate, completed pyrimidine nucleotides and a two-fold higher ether-linked to diacyl phosphatidylcholine ratio. The glutathione, NAD and NADP redox ratios were higher in AGS, whereas the S-adenosylmethionine to S-adenosylhomocysteine ratio was 24.5-fold higher in HGC-27. Each conditioned medium was compared with the fresh medium in which that line had been grown, identifying 27 metabolites whose exchange distinguished the two lines. Conclusions AGS and HGC-27 occupy two internally coherent metabolic states, differing at the triose phosphate branch of glycolysis and its glycerolipid product, at both ends of the mevalonate pathway, in the oxidation state of their redox couples and their membrane ether-lipid content, in which nucleotide branch is carried to completion, and in methylation index. Each of these pairings defines a specific functional test, from ferroptosis sensitivity to stable-isotope labelling of the glycerolipid and nucleotide branches. Keywords: Gastric cancer; Metabolomics; Lipidomics; AGS; HGC-27; Ether lipids