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  • Exometabolom analysis of breast cancer cell lines: Metabolic signature.

Exometabolom analysis of breast cancer cell lines: Metabolic signature.

Scientific reports (2015-08-22)
Lucas Willmann, Thalia Erbes, Sebastian Halbach, Tilman Brummer, Markus Jäger, Marc Hirschfeld, Tanja Fehm, Hans Neubauer, Elmar Stickeler, Bernd Kammerer
초록

Cancer cells show characteristic effects on cellular turnover and DNA/RNA modifications leading to elevated levels of excreted modified nucleosides. We investigated the molecular signature of different subtypes of breast cancer cell lines and the breast epithelial cell line MCF-10A. Prepurification of cell culture supernatants was performed by cis-diol specific affinity chromatography using boronate-derivatized polyacrylamide gel. Samples were analyzed by application of reversed phase chromatography coupled to a triple quadrupole mass spectrometer. Collectively, we determined 23 compounds from RNA metabolism, two from purine metabolism, five from polyamine/methionine cycle, one from histidine metabolism and two from nicotinate and nicotinamide metabolism. We observed major differences of metabolite excretion pattern between the breast cancer cell lines and MCF-10A, just as well as between the different breast cancer cell lines themselves. Differences in metabolite excretion resulting from cancerous metabolism can be integrated into altered processes on the cellular level. Modified nucleosides have great potential as biomarkers in due consideration of the heterogeneity of breast cancer that is reflected by the different molecular subtypes of breast cancer. Our data suggests that the metabolic signature of breast cancer cell lines might be a more subtype-specific tool to predict breast cancer, rather than a universal approach.

MATERIALS
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Guanosine, ≥97.0% (HPLC)
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Adenosine
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Ethylenediaminetetraacetic acid, 99.995% trace metals basis
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Ethylenediaminetetraacetic acid solution, 0.02% in DPBS (0.5 mM), sterile-filtered, BioReagent, suitable for cell culture
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Guanosine, BioReagent, suitable for cell culture
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Ethylenediaminetetraacetic acid, purified grade, ≥98.5%, powder
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Ethylenediaminetetraacetic acid, anhydrous, BioUltra, ≥99% (titration)
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Ethylenediaminetetraacetic acid, ACS reagent, 99.4-100.6%, powder
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Adenosine, ≥99%
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Adenosine, suitable for cell culture, BioReagent
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Ethylenediaminetetraacetic acid, anhydrous, crystalline, BioReagent, suitable for cell culture
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Xanthosine dihydrate, ≥99%
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Inosine, ≥99% (HPLC)
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Methanol solution, contains 0.10 % (v/v) formic acid, UHPLC, suitable for mass spectrometry (MS), ≥99.5%
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HEPES buffer solution, 1 M in H2O
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Formic acid, ≥95%, FCC, FG
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Guanosine, ≥98%
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Methanol solution, NMR reference standard, 4% in methanol-d4 (99.8 atom % D), NMR tube size 3 mm × 8 in.
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Trypan Blue solution, 0.4%, liquid, sterile-filtered, suitable for cell culture
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Methanol-12C, 99.95 atom % 12C
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S-(5′-Adenosyl)-L-homocysteine, crystalline
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Sodium chloride solution, 5 M
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Methyl-β-D-thiogalactoside
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HEPES, ≥99.5% (titration)
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HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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Sodium chloride, BioXtra, ≥99.5% (AT)
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HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
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5′-Deoxy-5′-(methylthio)adenosine
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Hydrocortisone, BioReagent, suitable for cell culture
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Uridine, BioUltra, ≥99%