快速筛选雌激素类环境激素的方法探讨

桂黎明; 任宏伟; 杨功名

快速筛选雌激素类环境激素的方法探讨

Rapid and sensitive screening method for detecting estrogenic compounds in environmental hormone

摘要

摘要:
目的开发快速筛选环境激素的检测方法.
方法将构建的表达人α雌激素受体的载体pcDNA3.1-ER和含雌激素受体反应元件的荧光素酶报告基因载体pGL3-tk-ERE共转染到HeLa细胞中, 以雌二醇或被检样品双酚A诱导雌激素受体表达后检测荧光素酶活性.
结果报告基因载体中的荧光素酶的表达受雌激素受体反应元件的调控, 且在一定浓度范围内荧光素酶的活性与雌二醇的加入量成线性关系; 疑为有雌激素样作用的双酚A也显著诱导荧光素酶基因的表达.
结论本方法经济、简便、快速, 有望用于快速筛选疑有雌激素样作用的化学物质.

Abstract:
Objective To investigate a rapid and sensitive scr eening method for detecting estrogenic compounds in enviromnental hormone.
Methods Both expression plasmid pcDNA31.1-ER and repor ter plasmid pGL3-tk-ERE were constructed by inserting cDNA of human estrogen receptor-α(ER-α)into plasmid pcDNA31.1 and estrogen receptor response element(ERE)into reporter plasmid pGL3-tk, respectively.After cotransfection the two plasmids into Hela cells, the luciferase activity induced by 17-β-estradiol(E₂)or Bisphenol A(BPA)through the interaction of ER with ERE were detected.
Results the activity of enzyme regulated by ERE in the presence of E₂ and BPA were linearly correlated with the concentration of E₂ and BPA within a certain dose range.
Conclusion this method was suitable for detection of estrogen and estrogenic compounds.With further development, it would become a rapid and efficient routine method for detection of estrogenic compounds in the environmental hormone.

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