基于报告基因的化学预防剂细胞筛选模型的建立  

Establishment of reporter gene-based cell screening model for discovering new chemopreventive agents

作　　者：徐海荣[1] 卜平[1] 李湘鸣[1] 

机构地区：[1]扬州大学医学院中西医结合研究所,江苏扬州225001

出　　处：《中国中药杂志》2009年第14期1835-1839,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30873343);江苏省研究生培养创新工程项目(CX07S-042z)

摘　　要：目的:建立基于报告基因和抗氧化反应元件(anti-oxidative response element,ARE)的药物筛选细胞模型,为化学预防剂的筛选奠定基础。方法:构建重组报告基因表达载体p4ARE-TK-GFP/neo,并将其转入HepG2细胞中,利用已知化学预防剂检测该细胞模型,并观察白藜芦醇、原儿茶醛、熊果酸和齐墩果酸4种中药单体不同浓度(0,12.5,25,50,100,200μmol.L-1)对GFP报告基因表达活性的影响。结果:表达载体p4ARE-TK-GFP/neo中GFP的表达受ARE的调控,其表达水平高于对照载体且在一定范围内与化学预防剂的浓度呈剂量效应关系。4种中药单体中白藜芦醇有较好的诱导效果。结论:利用此模型通过测定GFP报告基因的诱导表达水平可筛选得到新结构的化学预防剂。Objective： To discover new chemopreveutive agents, a drug screening cell model based on reporter gene and anti- oxidative response element （ARE） has been established. Method ： Four repeas of ARE DNA binding conserved sequences were syn- thesized 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 dif- ferent concentration （0, 12. 5, 25, 50, 100,200μmol.L-1 ） were observed by determining reporter gene GFP activity. Result： 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 res- veratrol was significantly increased. Conclusion： The method can be used to screening of chemopreventive agents from Chinese tradi- tional medicine by measurement of luminescent value of expressed GFP in wells of microtiter plate.

关 键 词：抗氧化反应元件 报告基因 化学预防剂 中药 药物筛选模型 

分 类 号：R96[医药卫生—药理学]