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目的 建立基于报告基因的雌激素受体α亚型配体的筛选模型 ,用此模型对收集到的化合物进行筛选 ,以发现具有新结构的雌激素受体配体。方法 构建诱导性表达载体并转染入ERα +HepG2细胞中 ,筛选报告基因CAT表达受雌二醇诱导的阳性克隆。采用ELISA法检测化合物对CAT表达的影响 ,体外细胞存活实验对化合物进行功能性检测。结果 ERα +HepG2细胞中CAT的表达受雌二醇的诱导并呈现剂量依赖关系。化合物三羟基二苯乙烯诱导CAT表达的最大活性为雌二醇的 1 75倍。结论 利用此模型通过测定CAT基因的诱导表达水平可筛选得到新结构的雌激素受体配体。
OBJECTIVE: To establish a screening model of reporter gene-based estrogen receptor alpha subtype ligands and screen the collected compounds with this model to find estrogen receptor ligands with novel structures. Methods The recombinant plasmid was constructed and transfected into ERα + HepG2 cells. The positive clones whose expression of CAT gene was induced by estradiol were screened. The effect of compound on the expression of CAT was detected by ELISA, and the cell viability was tested in vitro. Results The expression of CAT in ERα + HepG2 cells was induced by estradiol in a dose-dependent manner. The highest activity of compound trihydroxystilbene-induced CAT expression was 17O times that of estradiol. Conclusions The new structure of estrogen receptor ligand can be screened by measuring the expression level of CAT gene using this model.