The response of electrochemical method to estrogen effect and the tolerance to culture factors: Comparison with MTT and cell counting methods

Estrogen substances in the environment are increasing dramatically, which interfere with the normal hormone level of human body, lead to the disorder of endocrine system and even cancer. It is difficult to screen a large number of environmental estrogen substances by existing estrogen effect detecti...

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Veröffentlicht in:Analytica chimica acta 2022-11, Vol.1233, p.340514-340514, Article 340514
Hauptverfasser: Wei, Ying, Gao, Changsheng, Cui, Jiwen, Shen, Hongkuan, Zhao, Yanli, Zhou, Shi, Ye, Cai, Du, Yuan, Li, Jinlian, Wu, Dongmei
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container_start_page 340514
container_title Analytica chimica acta
container_volume 1233
creator Wei, Ying
Gao, Changsheng
Cui, Jiwen
Shen, Hongkuan
Zhao, Yanli
Zhou, Shi
Ye, Cai
Du, Yuan
Li, Jinlian
Wu, Dongmei
description Estrogen substances in the environment are increasing dramatically, which interfere with the normal hormone level of human body, lead to the disorder of endocrine system and even cancer. It is difficult to screen a large number of environmental estrogen substances by existing estrogen effect detection methods, and the results are often affected by many factors, thus the development of new method has become an urgent task. Electrochemical method is promising to reflect cell proliferation by tracking intracellular purine bases directly. In this study, the estrogen level in MCF-7 cells on multiwall carbon nanotubes modified glassy carbon electrode (MWCNTs/GCE) could be tracked simply and conveniently, and the estrogen effect of estradiol could be reflected by electrochemistry in time and dose-dependent manners. Electrochemical method displayed the best tolerance to culture factors, such as different cell densities, serum types, culture medium types and serum estrogen-free methods, which responsed to estrogen effect higher than MTT (about 40%) and cell counting methods (about 50%). Further Western blotting analysis showed that the estrogen effect of estradiol promoted purine catabolism and up-regulated guanine deaminase (GDA) and adenine deaminase (ADA) expression, the key enzymes of purine catabolism pathway, in a dose-dependent manner. The up-regulation of GDA and ADA led to the increase of intracellular guanine and xanthine, which enhanced the electrochemical signal derived from guanine and xanthine. [Display omitted] •Electrochemistry could track estrogen level in MCF-7 cells simply and conveniently.•Electrochemistry displayed excellent environment tolerance compared to other methods.•E2 estrogen effect up-regulated GDA and ADA expression in purine catabolism pathway.
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Further Western blotting analysis showed that the estrogen effect of estradiol promoted purine catabolism and up-regulated guanine deaminase (GDA) and adenine deaminase (ADA) expression, the key enzymes of purine catabolism pathway, in a dose-dependent manner. The up-regulation of GDA and ADA led to the increase of intracellular guanine and xanthine, which enhanced the electrochemical signal derived from guanine and xanthine. 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subjects Electrochemistry
Estradiol
Estrogen effect
Purine catabolism
title The response of electrochemical method to estrogen effect and the tolerance to culture factors: Comparison with MTT and cell counting methods
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