Aim To establish a method of drug screening based on
reporter gene and the signal transduction of PPARγ(Peroxisome proliferator-activated
receptor γ) system for discovering the small molecular compounds with activity of improving insulin sensitivity.Methods A recombinant vector PPRE-tk-Luc~+ was transiently cotransfected with a PPARγ expression vector pCMX- PPARγ into five cell lines respectively. The effect of
rosiglitazone on the signal transduction of PPARγ was determined by examing the luciferase activity, the cell line with the highest induction rate was selected for further experiments. The speciality of induction was examined by the ligands of nuclear receptors.Results The expression levels of
reporter gene induced by rosiglitazone in 293T cells was the highest, up to 4.9 fold, and in dose dependent manner. The Z'factor was 0.72. The expression levels of luciferase in 293T cells induced by the ligands of nuclear receptors were about 1.0 fold. No effect of rosiglitazone on the proliferation of 293T was observed.Conclusions The expression of luciferase in 293T cells co-transfected with the recombinant vector PPRE-tk-Luc~+ and pCMX-PPARγ vector could be induced significantly. The preferable speciality and stability of the cellular model make it well suited for establishing high-throughput screening systems for agonists of PPARγ.