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Breast Cancer Research and Treatment

, Volume 131, Issue 1, pp 49–63 | Cite as

Glucocorticoid receptor activity discriminates between progesterone and medroxyprogesterone acetate effects in breast cells

  • Aurélie Courtin
  • Laudine Communal
  • Myriam Vilasco
  • Daniela Cimino
  • Najat Mourra
  • Michele de Bortoli
  • Daniela Taverna
  • Anne-Marie Faussat
  • Marc Chaouat
  • Patricia Forgez
  • Anne Gompel
Preclinical study

Abstract

The purpose of this article is to determine the tumorigenic potential of estradiol treatment (E2) when combined with either progesterone (P4) or medroxyprogesterone acetate (MPA) in normal luminal human breast cells (HBE) and in human breast cancer cells (T47-D, MCF-7). Proliferation profiles were evaluated, along with the gene transactivation activity between the progesterone and glucocorticoid receptors (PR, GR) in HBE, T47-D, and MCF-7 cells treated by E2 + P4 or E2 + MPA. High throughput transcriptome analysis was performed on RNA from HBE cells treated by E2, E2 + MPA and E2 + P4. GR content was analyzed in normal breast cells as well. In HBE cells, E2 + P4 treatment was antiproliferative and promoted cellular differentiation. In contrast, E2 + MPA displayed mitogenic, antiapoptotic effects in HBE cells and did not influence cellular differentiation. The effect of P4 and MPA on cell proliferation was, however, variable in breast cancer cells. In cells containing GR or/and PR, MPA decreased proliferation whereas P4 antiproliferative effect needed the presence of PR. In HBE cells, the regulation of genes by E2 + P4, and E2 + MPA was significantly different, particularly in cell proliferation and cell death gene families. Further analysis revealed a modulation of the glucocorticoid receptor gene expression pathway by E2 + MPA. Predominant MPA glucocorticoid activity in normal and breast cancer cells was demonstrated using a glucocorticoid antagonist and the down-regulation of the GR by RNA interference. In normal luminal breast cells and in breast cancer cells, P4 and MPA combined with E2 treatment have opposing mitogenic effects due to GR. The consequences of MPA glucocorticoid potencies as well as the importance of GR in breast tissue merit a reappraisal.

Keywords

Proliferation Differentiation Microarrays Normal breast cells Glucorticoid Glucocorticoid receptor silencing Antiglucocorticoid Antiprogestin VA-2914 FAS IEX-1 

Notes

Acknowledgments

We thank Drs. Michèle Resche-Rigon (HRA Pharma, Paris) for helpful discussions and providing the anti-progestogen VA-2914, Steve K. Nordeen (University of Colorado, Denver) for kindly providing sequences of IEX-1 primers, Helenius J. Kloosterboer (Organon, Part of Shering-Plough, Oss) for providing antiglucocorticoid ORG-34116, Ramiro Castro Urology Medical Director (GSK, UK) for the generous gift of Dutasteride and Sylvie Dumont for her excellent technical assistance in immunohistochemistry experiments. We are grateful to Niel Insdorf for his kind help in editing the manuscript. This research was supported by grants from INSERM-UPMC, the Association pour la Recherche sur le Cancer, HRA Pharma and Novartis (France). Aurélie Courtin was a recipient of a grant from the Association pour la Recherche sur le Cancer, Laudine Communal is a recipient for a grant from HRA Pharma. Myriam Vilasco is a recipient of a post-doc fellowship (Institut National Contre le Cancer-INCA). Daniela Taverna was supported by grants from the University of Torino (Local Research Funding 2007/DT, 2008/DT), Regione Piemonte Ricerca Scientifica Applicata (CIPE2004/DT) and Compagnia di San Paolo, Torino. Daniela Cimino is a fellow of the Regione Piemonte.

Supplementary material

10549_2011_1394_MOESM1_ESM.ppt (1.3 mb)
Supplementary Figure 1: Validation of HBE cell model: PR and ER protein expression. Immunocytochemistry of PR (left) and ER (right) and negative control (NC) (a) (original magnification ×200). Characterization of luminal epithelial cells using expression of CK18 detected by immunofluorescence (b) (original magnification ×600). Comparison of PR and ER mRNA amounts in normal and breast cancer cells by RT-qPCR (c). Results are expressed in cycle threshold (Ct) values (mean ± SEM, n = 5 (HBE), n = 3 (MCF-7 and T47-D))

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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Aurélie Courtin
    • 1
  • Laudine Communal
    • 1
  • Myriam Vilasco
    • 1
  • Daniela Cimino
    • 2
    • 3
  • Najat Mourra
    • 1
    • 4
  • Michele de Bortoli
    • 2
  • Daniela Taverna
    • 2
    • 3
  • Anne-Marie Faussat
    • 5
  • Marc Chaouat
    • 6
  • Patricia Forgez
    • 1
  • Anne Gompel
    • 1
    • 7
  1. 1.INSERM-UPMC, UMRS 938, Hôpital Saint-AntoineParisFrance
  2. 2.Center for Molecular Systems Biology and Department of Oncological SciencesUniversity of Torino, c/o Institute for Cancer Research and Treatment (IRCC)CandioloItaly
  3. 3.Molecular Biotechnology CenterUniversity of TorinoTurinItaly
  4. 4.Service d’Anatomie et Cytologie Pathologiques, AP-HP, Hôpital Saint-AntoineParisFrance
  5. 5.Plateforme de Cytométrie, IFR 65-St Antoine UMPCParisFarnce
  6. 6.Université Paris Diderot, AP-HP, Hôpital Saint-Louis, Service de Chirurgie PlastiqueParisFrance
  7. 7.Université Paris Descartes, AP-HP, Hôtel-Dieu, UF de GynécologieParisFrance

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