[논문]Assessment of the Estrogenicity of Isoflavonoids, Using MCF-7-ERE-Luc Cells

Joung, Ki-Eun; Kim, Yeo-Woon; Sheen, Yhun-Yhong

doi:10.1007/bf02976687

Assessment of the Estrogenicity of Isoflavonoids, Using MCF-7-ERE-Luc Cells 원문보기

Archives of pharmacal research : a publication of the Pharmaceutical Society of Korea, v.26 no.9, 2003년, pp.756 - 762

Joung, Ki-Eun (College of Pharmacy, Ewha Womans University) , Kim, Yeo-Woon (College of Pharmacy, Ewha Womans University) , Sheen, Yhun-Yhong (College of Pharmacy, Ewha Womans University)

Abstract ▼ AI-Helper

In the current study, our research focused on the estrogenic activity of isoflavonoids, mainly genistein, biochanin A and daidzein. Genistein enhanced the reporter gene expression of MCF-7-ERE-Luc cells, at a concentration as low as 10 nM, with a concentration of 100 nM the achieved gene expression effects were similar to those of 10 pM 17$\beta$-estradiol. Based on the estrogenic activities of biochanin A and daidzein, hydroxyl groups at the 4 and 5 positions are needed for the maximal effect of the genistein. The estrogenic effects of these isoflavonoids were inhibited by the concomitant treatment with tamoxifen. The data showed that the estrogenic effects of isoflavonoids were mediated through estrogen receptors. When the isoflavonoids were tested as mixtures, the estrogenic effects were lower than the arithmetic sum of those induced by each individual isoflavonoid. The estrogenic potency of each isoflavonoid was presented at EC50 levels with a 17$\beta$-estradiol equivalent concentration (EEQ) based on the dose response of each chemical. The EC50s and EEQs of genistein, biochanin A and daidzein were 4.15, 0.89 and 0.18 $\mu$M, and 15.0, 5.12 and 1.83 $\mu$ M/M, respectively. Our data clearly demonstrated that the pERE-luciferase reporter gene assay was suited for the sensitive and quantitative measurement, and large scale screening, of the estrogenicity of chemicals in vitro.

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제안 방법

Isoflavonoids were tested in combination, in order to determine the potential interactivating effects of exposure to these mixtures. As shown in Fig.
The MCF-7 cells were maintained in minus phenol red medium for 6 days prior to the 17p-estradiol treatment, were then treated with various concentrations of 17p-estradiol for 6 days, and the cell numbers measured based on the SRB assay. As the concentration of the 17p-estradiol increased, the numbers of cells increased, in a dose de pendent manner.
We, therefore, have 니sed the plasmid that contains the ERE upstream of the 山ciferase gene 4 stable transfection of this plasmid, into MCF-7 hurric n breast cancer cells, where estrogen receptors are abundant, was obtained. Using these MCF-7-ERE-Luc cells, wo have evaluated the relationship of the struct니re of iscfla /onoids and their estrogenic activities.

성능/효과

5A). A combination of 5 卩M genistein and 1 |1M biochanin A showed 160% of the activity of 10 pM 17p-estradiol, whereas biochanin A 기one had 95% of the activity of 10 pM 17p-estradiol, and 5 jiM genistein 기one had 190% of the activity of 10 pM 17p estradiol. Likewise, the combined estrogenic activity of genistein and biochanin A (160%) appeared to be lower than the arithmetic sum of the individual estrogenic activities (Fig.
0 for windows. The EC₅₀s of the 17p-es- tradiol and the test chemicals were calculated by deter mining the concentration at which 50% of the maximum luciferase activity was attained. The detection limit was calculated as the hjciferase activity elicited by the control, plus three times the standard deviation, To calculate CALUX EEQ, the luciferase response ca니sed by the test chemicals was interpolated over the linear range of the corresponding 17p-estradiol standard curve.

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Assessment of the Estrogenicity of Isoflavonoids, Using MCF-7-ERE-Luc Cells 원문보기

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Assessment of the Estrogenicity of Isoflavonoids, Using MCF-7-ERE-Luc Cells 원문보기

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