title: A Cellular Assay for Inhibitors of the Fatty Acid Biosynthetic Pathway Using Scintillating Microplates.
abstract: A simplified method for monitoring the incorporation of radiolabeled acetate into lipids in a cellular system is described. The assay eliminates the commonly employed labor-intensive organic extraction step by plating the cells in 96-well tissue culture-treated ScintiPlates(®) that enable direct measurement of radiolabeled cell membrane-embedded lipids. Since the scintillant is entrenched in the plates, radioactivity in close proximity to the scintillant is measured without the need for liquid scintillation cocktail. The utility of this method for evaluating inhibitors of the de novo fatty acid synthetic pathway is demonstrated here with fatty acid synthase (FASN). Due to the upregulation of FASN activity in many tumor types, development of inhibitors to block the FASN activity in cells shows promise as an attractive and tractable approach for therapeutic intervention.
authors.affiliation: 1 Lead Discovery and Profiling, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Thao
authors.initials: T
authors.lastname: Ung
authors.affiliation: 1 Lead Discovery and Profiling, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Mason
authors.affiliation: 1 Lead Discovery and Profiling, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Ron G
authors.initials: RG
authors.lastname: Robinson
authors.affiliation: 2 Global Bioassay and Technology, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Chrysanthe M
authors.initials: CM
authors.lastname: Spais
authors.affiliation: 1 Lead Discovery and Profiling, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Ator
authors.affiliation: 1 Lead Discovery and Profiling, Teva Branded Pharmaceutical Products R&D, Inc. , West Chester, Pennsylvania.
authors.forename: Thelma S
authors.initials: TS
authors.lastname: Angeles
mesh_terms: D000818:Animals; D053898:Biosynthetic Pathways / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D064429:Fatty Acid Synthases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005227:Fatty Acids / Q000037:antagonists & inhibitors / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D007313:Insecta; D012588:Scintillation Counting / Q000379:methods*
chemical_list: D004791:Enzyme Inhibitors; D005227:Fatty Acids; D064429:Fatty Acid Synthases