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 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.
Piccioni
Fabiana
F
Department of Chemistry, University of Rome La Sapienza, P.le A. Moro 5, 00185 Rome, Italy.
Borioni
Anna
A
Delfini
Maurizio
M
Del Giudice
Maria Rosaria
MR
Mustazza
Carlo
C
Rodomonte
Andrea
A
Risuleo
Gianfranco
G
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004121:Dimethyl Sulfoxide; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051379:Mice; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D009711:Nucleotides / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014508:Urea / Q000031:analogs & derivatives / Q000494:pharmacology
D000596:Amino Acids; D004791:Enzyme Inhibitors; D009711:Nucleotides; D011743:Pyrimidines; D014508:Urea; C496349:Fgfr1 protein, mouse; D051496:Receptor, Fibroblast Growth Factor, Type 1; C113399:PD 166866; D004121:Dimethyl Sulfoxide