title: A cellular assay for measuring the modulation of glucose production in H4IIE cells.
abstract: Type II diabetes and its associated complications are a major health concern of the developed world. One of the hallmarks of diabetes is insulin resistance, where secreted insulin no longer has any effect on its target tissues, namely, liver, muscle, and fat. An important therapeutic strategy is to modulate blood glucose levels using pharmacological agents. Glycogen synthase kinase-3 (GSK3) is a serine-threonine protein kinase that plays important roles in regulating glucose metabolism. It is a key negative regulator of insulin action and is an important contributing factor to insulin resistance in liver, muscle, and adipose tissue. We describe the development of a cell-based assay designed to measure glucose production in rat hepatoma cell line H4IIE liver cells in response to treatment with small molecule inhibitors, including GSK3 inhibitors. The assay is set up in a 96-well format, and glucose production is assessed using a convenient fluorescence-based readout. This disease-relevant cellular assay is a valuable tool for the progression of small molecules that modulate glucose production.
authors.affiliation: Serono Pharmaceutical Research Institute, Plan-les-Ouates, Switzerland.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: de Raemy-Schenk
authors.forename: Solenne
authors.initials: S
authors.lastname: Troublé
authors.forename: Pascale
authors.initials: P
authors.lastname: Gaillard
authors.forename: Patrick
authors.initials: P
authors.lastname: Page
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Gotteland
authors.forename: Alexander
authors.initials: A
authors.lastname: Scheer
authors.forename: Paul
authors.initials: P
authors.lastname: Lang
authors.forename: Karen
authors.initials: K
authors.lastname: Yeow
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D005947:Glucose / Q000378:metabolism*; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism*; D007328:Insulin / Q000008:administration & dosage*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008657:Metabolic Clearance Rate / Q000187:drug effects; D051381:Rats
chemical_list: D007328:Insulin; D038362:Glycogen Synthase Kinase 3; D005947:Glucose