评价环境雄激素类化合物重组Lac Z报告基因酵母细胞的建立  

Establishment of recombinant Lac Z reporter gene-transformed yeast cells for bioassay of androgen-like compounds in environment

作　　者：戴卫星[1] 杨晨[1] 罗方妮[2] 李姗姗[1] 黄仁华[1] 谢阳梅[1] 薛飞玥[1] 李湘鸣[1] 

机构地区：[1]扬州大学医学院预防医学教研室,江苏扬州225001 [2]扬州大学动物科学与技术学院,江苏扬州225009

出　　处：《细胞与分子免疫学杂志》2013年第3期303-309,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：江苏省环境监测科研基金(0714);2011年江苏省大学生实践创新训练计划(批准号B11129);扬州大学2011年度大学生学术科技创新基金(33)

摘　　要：目的建立环境雄激素内分泌干扰物酵母评价体系。方法以载体双表达的方式构建该重组基因酵母细胞。在表达载体中,用3-磷酸甘油醛脱氢酶(GPD)启动子驱动雄激素受体基因(AR)的表达,并使其与V5抗原表位相融合;在报告载体中,用雄激素效应元件(ERE)调控的Lac Z作为报告基因。将两者转化于酵母细胞(W303-1A)中,构建成雄激素调控的重组Lac Z基因酵母细胞。用不同浓度的雄激素[去氢表雄酮(DHT)和丙酸睾酮(TP)]和雌激素(雌二醇、雌三醇、雌酮、二乙基已烯雌酚和乙炔基雌二醇),对重组基因酵母细胞分别进行敏感度和特异度研究。结果去氢表雄酮和丙酸睾酮与该重组基因酵母细胞有明显的剂量效应关系,说明其具有良好的特异度,而与雌二醇、雌三醇、雌酮、二乙基已烯雌酚和乙炔基雌二醇5种雌激素化合物均无明显的剂量效应关系,说明其具有较强的特异性。结论该重组基因酵母细胞可用于对环境雄激素类化合物的筛选。Objective To establish the yeast-based bioassay system for the androgenic endocrine disruptors in the envi- ronment. Methods The recombinant gene-transformed yeast cells were constructed based on two episomal vectors. In the expression vector, the expression of androgen receptor （AR） gene was driven by 3-glyceraldehydephosphate dehydrogen- ase （GPD） promoter and the gene was fused to V5 epitope; in the reporter vector, the expression of the Lac Z gene as a report gene was under the control of the androgen response element （ARE）. The vectors were transformed into yeast cells （W303-1A） to construct the yeast-based bioassay system. The sensitivity of this assay system was evaluated by the andro- genic compounds （dihydrotestosterone, DHT and testosterone propionate, TP）and the specificity of this system by the estro- genic compounds （ 1715-estradiol, estriol, estrone, diethylstilbestrol and ethinyl estradiol）. Results Both DHT and TP yielded a significant dose-effect model with the recombinant gene-transformed yeast cells, indicating that this yeast bioassay system has a high sensitivity. Moreover, the assay also exhibited a high specificity in that the yeast cells had no dose-effect relation- ship with the different concentrations of five estrogens （1713-estradiol, estriol, estrone, diethylstilbestrol and ethinyl estradiol）. Conclusion The recombinant yeast cells could be applied to screen for the androgen-like compounds in the environment.

关 键 词：酵母细胞 雄激素受体 雄性内分泌干扰物 

分 类 号：R392.12[医药卫生—免疫学] R122.11[医药卫生—基础医学]