Effect of insulin, cAMP, and dexamethasone on phosphoenolpyruvate carboxykinase gene promoter in vitro.
- Author:
Kai FENG
1
;
Heng WANG
;
Qi SUN
;
Xin-Hua XIAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Line, Tumor; Cyclic AMP; pharmacology; Dexamethasone; pharmacology; Dose-Response Relationship, Drug; Gene Expression Regulation, Enzymologic; drug effects; Insulin; administration & dosage; pharmacology; Liver Neoplasms, Experimental; pathology; Luciferases; genetics; metabolism; Phosphoenolpyruvate Carboxykinase (GTP); genetics; metabolism; Promoter Regions, Genetic; drug effects; genetics; Rats; Recombinant Proteins; genetics; metabolism; Transfection
- From: Acta Academiae Medicinae Sinicae 2004;26(6):639-642
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of insulin, cyclic adenosine monophosphate (cAMP), and dexamethasone (DEX) on 550 bp (-600 -/+ 69) fragment of phosphoenolpyruvate carboxykinase (PEPCK) gene promoter by reporter gene.
METHODSThe recombinant pGL2-PEPCK-Luc and the control plasmid pSV-beta-Galactosidase were co-transfected to rat hepatoma cell line (CBRH7919) by lipofectin. By measuring luciferase activity, we evaluated in vitro regulation of PEPCK gene promoter on reporter gene transcription.
RESULTScAMP and DEX stimulated PEPCK promoter obviously; meanwhile, they also had accumulative effects. At different physiological concentrations, insulin had a suppressive effect on PEPCK promoter, which was dose-independent.
CONCLUSIONThere is a perfect feedback mechanism for PEPCK promoter in hepatoma cell. 550 bp (-600 -/+ 69) fragment of PEPCK may be a candidate gene in the gene therapy of diabetes.
