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Anti-estrogenic mechanism of unliganded progesterone receptor isoform B in breast cancer cells

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Abstract

Over half of breast cancer cases are estrogen-dependent and strategies to combat estrogen-dependent breast cancer have been to either block the activation of estrogen receptor (ER) or diminish the supply of estrogens. Our previous work documented that estrogen-independent expression of progesterone receptor (PR) in MCF-7 cells markedly disrupted the effects of estrogen. In this study, we have developed an adenovirus-mediated gene delivery system to study the specific involvement of PR isoform A (PR-A) and PR-B in the anti-estrogenic effect and its mechanism of action. The results revealed that PR-B, but not PR-A, exhibited distinct anti-estrogenic effect on E2-induced cell growth, gene expression, and ER-ERE interaction in a ligand-independent manner. The anti-estrogenic effect of PR-B was also associated with heightened metabolism and increased cellular uptake of estradiol-17β (E2). We have also found that the B-upstream segment of PR-B alone was able to inhibit E2-induced ER-ERE interaction and cellular uptake of E2. Although PR-A alone did not affect E2-induced ER activity, it antagonized the anti-estrogenic effect of PR-B in a concentration-dependent manner. The findings suggest an important mechanism of maintaining a favorable level of ER activity by PR-A and PR-B in estrogen target cells for optimal growth and differentiation. The potential anti-estrogenic mechanism of PR-B may be exploited for breast cancer therapy.

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Abbreviations

E2:

Estradiol-17β

ER:

Estrogen receptor

ERE:

Estrogen response element

PR:

Progesterone receptor

PR-A:

PR isoform A

PR-B:

PR isoform B

Ad/Φ:

Control adenovirus

Ad/PRA:

Adenovirus carrying PR-A cDNA

Ad/PRB:

Adenovirus carrying PR-B cDNA

MCF-7-Ad/Φ:

MCF-7 cells infected with Ad/Φ

MCF-7-Ad/PRA:

MCF-7 cells infected with Ad/PRA

MCF-7-Ad/PRB:

MCF-7 cells infected with Ad/PRB

BUS:

B upstream segment

AF3:

Activation function 3

DBD:

DNA binding domain

VR:

Variable region

NLS:

Nuclear location signal

HPLC:

High performance liquid chromatography

aa:

Amino acid

MOI:

Multiplicity of infection

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Acknowledgments

We thank Prof. Pierre Chambon from the Institute of Genetics and Molecular and Cellular Biology, Strasbourg, France, for providing the expression vectors hPR1 and hPR2, and Prof. Benita S. Katzenellenbogen, University of Illinois at Urbana-Champaign, Illinois, for providing the promoter interference constructs. This work is supported by Ministry of Education, Republic of Singapore (ARC/02) to Valerie C-L. Lin.

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Authors and Affiliations

  1. School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore

    Ze-Yi Zheng, Si-Min Zheng & Valerie C-L Lin

  2. Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, 4 Medical Drive, Singapore, 117597, Singapore

    Ze-Yi Zheng & Boon-Huat Bay

  3. Department of Clinical Research, Singapore General Hospital, Outram Road, Singapore, 169608, Singapore

    Ze-Yi Zheng & Swee-Eng Aw

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  1. Ze-Yi Zheng
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Correspondence to Valerie C-L Lin.

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Zheng, ZY., Zheng, SM., Bay, BH. et al. Anti-estrogenic mechanism of unliganded progesterone receptor isoform B in breast cancer cells. Breast Cancer Res Treat 110, 111–125 (2008). https://doi.org/10.1007/s10549-007-9711-8

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