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DOI: 10.1055/s-0028-1088817
Metabolic signature of breast cancer cell line MCF–7: Profiling of modified nucleosides via LC-IT MS coupling
Cancer is accompanied with metabolic disorders and shows characteristic effects on the activity of modifiying enzymes and DNA/RNA modifications, resulting in elevated amounts of excreted modified nucleosides. To understand the impaired RNA metabolism in breast cancer, we screened these metabolites the cell culture supernatants of the breast cancer cell line MCF–7 and compared it to the human mammary epithelial cells MCF–10A. The nucleosides were isolated and analyzed via 2D-chromatographic techniques: In the first dimension by cis-diol specific boronate affinity chromatography and subsequently by reversed phase chromatography coupled to an ion trap mass spectrometer. In total, 36 metabolites were identified by comparison of fragmentation patterns and retention time. Relation to the internal standard isoguanosine yielded normalized area ratios of each identified compound and enabled a semi-quantitative metabolic signature of both analyzed cell lines. 13 of the identified 26 modified ribonucleosides were elevated in the cell culture supernatants of MCF–7 cells, with 5-methyluridine, N2,N2,7-trimethylguanosine, N6-methyl-N6-threonylcarbamoyladenosine and 3-(3-aminocarboxypropyl)-uridine showing the most significant differences. 1-ribosylimidazole–4-acetic acid, a histamine metabolite, was solely found in the supernatants of MCF–10A cells, whereas 1-ribosyl–4-carboxamido–5-aminoimidazole and S-adenosylmethionine occurred only in supernatants of MCF–7 cells. The results are discussed against the background of pathological changes in cell metabolism, resulting in new perspectives for modified nucleosides and related metabolites as possible biomedical markers for breast carcinoma in vivo.
metabolomics - nucleosides