Metabolic alterations in cultured mouse fibroblasts induced by an inhibitor of the tyrosine kinase receptor Fibroblast Growth Factor Receptor 1

Proton nuclear magnetic resonance (NMR) spectroscopy was used to identify and quantify the metabolites present in cultured mouse fibroblast cells 3T6 in their native state and after treatment with PD166866, an inhibitor of the fibroblast growth factor receptor. Cell extracts were prepared according...

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Bibliographic Details
Published in:Analytical biochemistry 2007-08, Vol.367 (1), p.111-121
Main Authors: Piccioni, Fabiana, Borioni, Anna, Delfini, Maurizio, Giudice, Maria Rosaria Del, Mustazza, Carlo, Rodomonte, Andrea, Risuleo, Gianfranco
Format: Article
Language:English
Subjects:
Amino Acids - metabolism
Animals
Cell Line
Cultured murine fibroblasts
Dimethyl Sulfoxide
Enzyme Inhibitors - pharmacology
In vitro
Metabolic profiling
Mice
Nuclear Magnetic Resonance, Biomolecular - methods
Nucleotides - metabolism
Proton NMR spectroscopy
Pyrimidines - pharmacology
Receptor, Fibroblast Growth Factor, Type 1 - antagonists & inhibitors
Receptor, Fibroblast Growth Factor, Type 1 - metabolism
Urea - analogs & derivatives
Urea - pharmacology
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Summary:Proton nuclear magnetic resonance (NMR) spectroscopy was used to identify and quantify the metabolites present in cultured mouse fibroblast cells 3T6 in their native state and after treatment with PD166866, an inhibitor of the fibroblast growth factor receptor. Cell extracts were prepared according to the Bligh–Dyer protocol which prevents artifacts deriving from the chemical demolition of macromolecules. Also the growth medium was subjected to the same extraction procedure. The NMR approach made possible the identification and quantification of about 40 different metabolites at nanomoles/mg of protein level: the biological relevance of the variation of some metabolite levels is discussed. Our experimental procedure offers a prospective method for the evaluation of variations of the metabolic profile deriving from different biochemical treatments of these cells.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2007.04.013