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MVLN细胞:一种源自MCE - 7的生物发光细胞系,用于研究雌激素活性的调节。

MVLN cells: a bioluminescent MCE-7-derived cell line to study the modulation of estrogenic activity.

作者信息

Demirpence E, Duchesne M J, Badia E, Gagne D, Pons M

机构信息

INSERM U 58, Montpellier, France.

出版信息

J Steroid Biochem Mol Biol. 1993 Sep;46(3):355-64. doi: 10.1016/0960-0760(93)90225-l.

Abstract

We previously established a stable expression system in MCF-7 cells for the detection of (anti)estrogenic activity by assaying the reporter enzyme activity of firefly luciferase. In this cell line (called MVLN), the bioluminescent response can be measured either in the cellular homogenate, or in intact living cells. Here we present various potential experimental uses of this cellular model. First, we used this cell line to screen natural or synthetic molecules classified as full or partial (anti)estrogens and observed that their behavior towards our model was identical to that expected. Moreover, the bioluminescent response was in agreement with the natural responses like cellular proliferation or stimulation of the progesterone receptor. We then demonstrated the inhibitory effects of retinoic acid and 1,25 dihydroxyvitamin D3, two molecules which do not compete with estradiol for its receptor. We thus deduced that with this cell line an "antiestrogenic" effect which occurred at any step of the estrogenic action, might be detected. Finally, we showed that detection of luciferase activity in intact living cells was particularly helpful for investigating the evolution of estrogenic activity. For instance, we observed that long-term treatment of MVLN cells with an antiestrogen irreversibly decreased the bioluminescent response by more than 90%. This phenomenon affected all cells equally and could not be reversed, even by long-term estradiol treatment. We therefore conclude that this chimeric response faithfully reflects estrogenic action in the cell and can be used to develop different aspects of the endocrine research.

摘要

我们之前在MCF-7细胞中建立了一个稳定的表达系统,通过检测萤火虫荧光素酶的报告酶活性来检测(抗)雌激素活性。在这个细胞系(称为MVLN)中,可以在细胞匀浆或完整的活细胞中测量生物发光反应。在此,我们展示了这个细胞模型的各种潜在实验用途。首先,我们使用这个细胞系筛选被归类为完全或部分(抗)雌激素的天然或合成分子,观察到它们在我们的模型中的行为与预期一致。此外,生物发光反应与细胞增殖或孕酮受体刺激等天然反应一致。然后,我们证明了视黄酸和1,25-二羟基维生素D3的抑制作用,这两种分子不与雌二醇竞争其受体。因此,我们推断使用这个细胞系可以检测在雌激素作用的任何步骤发生的“抗雌激素”效应。最后,我们表明在完整活细胞中检测荧光素酶活性对于研究雌激素活性的演变特别有帮助。例如,我们观察到用抗雌激素长期处理MVLN细胞会使生物发光反应不可逆地降低超过90%。这种现象对所有细胞的影响是均等的,即使长期用雌二醇处理也无法逆转。因此,我们得出结论,这种嵌合反应忠实地反映了细胞中的雌激素作用,可用于开展内分泌研究的不同方面。

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