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Method for rapidly evaluating action mechanism and complex effect of endocrine disrupting compounds

A technology of endocrine disruptors and mechanism of action, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as large amounts of data, long experiment cycle, and inability to meet large-scale rapid evaluation, and achieve a low data volume Effect

Inactive Publication Date: 2013-08-28
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the construction of these models requires a large amount of data, the experiment period is long, and the cost is high, which cannot meet the large-scale rapid evaluation, especially the compound effect research close to the actual concentration of the environment.

Method used

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  • Method for rapidly evaluating action mechanism and complex effect of endocrine disrupting compounds
  • Method for rapidly evaluating action mechanism and complex effect of endocrine disrupting compounds
  • Method for rapidly evaluating action mechanism and complex effect of endocrine disrupting compounds

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Embodiment Construction

[0018] The present invention will be described in detail below in conjunction with specific embodiments. The following examples will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form.

[0019] Test drug: 17β-estradiol (E 2 ), o, p'-DDT were purchased from Sigma Company of the United States.

[0020] Preparation method: the standard product is dissolved in ethanol to make 1×10 -3 mol / L mother liquor and stored at -20°C, diluted with ultrapure water to the desired concentration before the experiment.

[0021] Model organism: human breast cancer MCF-7 cells.

[0022] Reagents: Total RNA Extraction Kit, Reverse Transcription Kit, RT-PCR Kit were purchased from Treasure Bioengineering (Dalian) Co., Ltd.

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Abstract

The invention discloses a method for rapidly evaluating action mechanism and complex effect of endocrine disrupting compounds. The method comprises the following steps of: cultivation of a human breast cancer MCF-7 cell; real-time fluorescent quantitative PCR (polymerase chain reaction) for the purpose of obtaining a dosage-effect curve of the endocrine disrupting compounds to be tested; mixing the endocrine disrupting compounds to be tested to form different concentration ratios of exposure liquid; lastingly exposing the cell for 24 hours; gene transcription level analysis by real-time fluorescent quantitative PCR after finishing exposing; collecting data to carry out factorial analysis of variance; drawing a marginal outline map; and evaluating the action mechanism and the complex effect of the endocrine disrupting compounds to be tested, wherein the endocrine disrupting compounds to be tested are 17beta-estradiol and o,p'-DDT. The complex effect of the endocrine disrupting compounds is evaluated by estrogen target genes in the human breast cancer MCF-7 cell on the basis of statistic analysis of data. The method is simple, fast and sensitive, and is a novel method for rapidly evaluating the action mechanism and the complex effect of the endocrine disrupting compounds.

Description

technical field [0001] The invention relates to environmental endocrine disruptors, in particular to a method for rapidly evaluating the action mechanism and compound effects of endocrine disruptors. Background technique [0002] In the actual environment, the coexistence of multiple environmental endocrine-disrupting chemicals (EDCs) makes the compound effect of EDCs arouse the great attention of researchers and regulatory agencies. In general, the concentration of EDCs in the environment is very low. However, toxicological studies have found that weaker estrogenic EDCs mixed at lower than their respective NOEC (no observed effect concentration) produced a very significant estrogenic effect. This requires us to face up to the compound effects of low concentrations of endocrine disruptors in the environment and develop simple and effective methods to evaluate the compound effects of endocrine disruptors. [0003] At present, the research on compound effect mainly uses two ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王鲁梅钟玲盈向霄叶璟张颖詹学佳
Owner SHANGHAI JIAO TONG UNIV