Establishment of a cell-based drug screening model for identifying agonists of human peroxisome proliferator-activated receptor gamma (PPAR?)

Objectives Peroxisome proliferator-activated receptor gamma (PPAR?) plays a critical role in regulation of diverse biological processes, including lipid metabolism and adipogenesis, cell division and apoptosis, and is involved in variety of disease conditions, such as obesity, atherosclerosis, inflammation and tumour. Developing a cell-based reporter gene model targeting PPAR? would be useful to screen human PPAR? agonists that could be beneficial to patients with these diseases. Methods We stably co-transfected human embryonic kidney (HEK) cell line 293T cells with phPPARg-IRES2-EGFP vector to express human PPARg (hPPARg), a reporter vector pPPRE 3-TK-LUC, and control vector pRL-CMV. The efficiency of the co-transfection was evaluated with flow cytometry of hPPARg expressing cells. Specificity of hPPARg activity was determined by dual luciferase reporter assay of co-transfected cells exposed to PPARg agonist rosiglitazone, PPARa agonist WY14643 and retinoic acid receptor alpha (RARa) agonist all-trans-retinoic acid (ATRA). Key findings The phPPARg-IRES2-EGFP co-transfected HEK293T cells showed concentration-and time-dependent luciferase induction upon exposure to the rosiglitazone, while WY14643 and ATRA were unable to activate the co-transfected HEK293T cells. Conclusions These data indicated that the HEK293T cells could be stably transfected with hPPARg. This cell-based drug screening platform could be used targeting specific nuclear receptor of hPPARg with effectiveness and specificity for hPPARg agonists discovery.