Establishment of reporter gene-based cell screening model for discovering new chemopreventive agents / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>To discover new chemopreventive agents, a drug screening cell model based on reporter gene and antioxidative response element (ARE) has been established.</p><p><b>METHOD</b>Four repeas of ARE DNA binding conserved sequences were synthesized and cloned into a GFP expression vector. This construct was stably transfected into HepG2 cells in vitro. The cell model was tested with known chemopreventive agents and the effects of resveratrol, protocatechuic aldehyde, ursolic acid and oleanolic acid at different concentration (0, 12.5, 25, 50, 100, 200 micromol x L(-1)) were observed by determining reporter gene GFP activity.</p><p><b>RESULT</b>The induce level of GFP was regulated by ARE and the dose-dependence in a certain range was observed. The induce level of GFP by resveratrol was significantly increased.</p><p><b>CONCLUSION</b>The method can be used to screening of chemopreventive agents from Chinese traditional medicine by measurement of luminescent value of expressed GFP in wells of microtiter plate.</p>