Evaluation of Estrogenic and Anti-estrogenic Activity of Endocrine Disruptors Using Breast Cancer Spheroids: a Comparative Study of T47D and MCF7 Cell Lines in 2D and 3D Models

Overview

Journal Front Toxicol

Date 2025 Mar 10

PMID 40061085

Authors

Katia Barbaro

Elisa Innocenzi

Valentina Monteleone

Daniele Marcoccia

Annalisa Altigeri

Alessia Zepparoni

Daniela Caciolo

Cristian Alimonti

Marta Mollari

Paola Ghisellini

Cristina Rando

Roberto Eggenhoffner

Maria Teresa Scicluna

Affiliations

Soon will be listed here.

Abstract

Introduction: The estrogenic and anti-estrogenic effects of endocrine disruptors were examined using two-dimensional 2D and three-dimensional 3D estrogen receptor-positive T47D and MCF7 human breast cancer cells.

Methods: The model system was used to test the plasticizer Bisphenol A (BPA), a known endocrine disruptor (EDs) with estrogen-like action, aga inst 17β-Estradiol (E2), the endogenous nuclear estrogen receptor (nERs) ligand, and the anti-estrogenic drug Fulvestrant (FUL). Spheroid formation and gene expression of estrogen-regulated markers (pS2 and TGFβ3) both in 2D and 3D cultures were used to establish the dose-dependent cellular effects of these substances, evaluated cell viability either by separately treating with the individual substances or in co-treatment.

Results: BPA exhibited a dose-dependent estrogenic activity in both 2D and 3D cultures, significantly influencing cell proliferation and gene expression of estrogen-regulated markers (pS2 and TGFβ3). In contrast, FUL displayed anti-estrogenic properties, effectively inhibiting the proliferative effects of E2, thereby highlighting the complex interactions between these compounds and the nERs pathways in human breast cancer cells.

Discussion: Our findings indicate that E2 and BPA significantly increase pS2 expression while decreasing TGFβ3, and that FUL co-treatment reverses these effects. Therefore, the model system could serve to observe the cell-mediated effects caused by the interaction of EDs with nERs. Through the use of these model systems - 2D and especially 3D, the latter of which allow better emulation of complex physiological and pathological processes occurring , the effects caused by EDs on nERs pathways can be detected and studied under various conditions. This approach performs as a preliminary screening tool to identify estrogenic substances, offering the potential to reduce reliance on experiments and contributing to improved environmental and health risk assessments.

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