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Hypermethylation of the progesterone receptor A in constitutive antiprogestin-resistant mouse mammary carcinomas

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Abstract

Most breast carcinomas that are estrogen receptor (ER) and progesterone receptor (PR) positive respond initially to an endocrine therapy, but over time, they develop resistance (acquired hormone resistance). Others, however, fail to respond from the beginning (constitutive resistance). Overcoming hormone resistance is one of the major desirable aims in breast cancer treatment. Using the medroxyprogesterone acetate (MPA)-induced breast cancer mouse model, we have previously demonstrated that antiprogestin-responsive tumors show a higher expression level of PR isoform A (PRA) than PR isoform B (PRB), while tumors with constitutive or acquired resistance show a higher expression level of PRB. The aim of this study was to investigate whether PRA silencing in resistant tumors was due to PRA methylation. The CpG islands located in the PRA promoter and the first exon were studied by methylation-specific PCR (MSP) in six different tumors: two antiprogestin-responsive, two constitutive-resistant, and two with acquired resistance. Only in constitutive-resistant tumors, PRA expression was silenced by DNA methylation. Next, we evaluated the effect of a demethylating agent, 5-aza-2′-deoxycytidine, on PRA expression and antiprogestin responsiveness. In constitutive-resistant tumors, 5-aza-2′-deoxycytidine treatment in vitro and in vivo restored PRA expression and antiprogestin RU-486 responsiveness. Furthermore, high levels of DNA methyltransferase (Dnmts) 1 and 3b were detected in these tumors. In conclusion, our results suggest that methyltransferase inhibitors in combination with antiprogestins may be effective in the treatment of constitutive-resistant carcinomas with a high DNA methyltransferase level.

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Abbreviations

5azadC:

5-Aza-2′-deoxycytidine

Dnmts:

DNA methyltransferases

E2 :

17-β-Estradiol

ER:

Estrogen receptor

ERα :

ER alpha

FCS:

Fetal calf serum

GR:

Glucocorticoid receptor

H&E:

Hematoxylin and eosin

HPF:

High power field

i.p.:

Intraperitoneal

M:

Methylated

MPA:

Medroxyprogesterone acetate

MSP:

Methylation-specific PCR

PI:

Propidium iodide

PR:

Progesterone receptor

PRA:

PR isoform A

PRB:

PR isoform B

RU-486:

Mifepristone

s.c.:

Subcutaneous

UM:

Unmethylated

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Acknowledgments

We are very grateful to Drs. Gorostiaga and Bolado for excellent technical support, to Dr. S. Vanzulli for her help in the histopathological evaluation of organs, to Laboratorios Craveri, Buenos Aires for providing MPA. VW received an award from Avon Foundation for data presented at the AACR Meeting 2009, the Breast Cancer Meeting 2009, and ICRETT Fellowship from the UICC permitting training in Dr Russo’s laboratory. This study was supported by Fundación Sales and SECyT (PICT05, 05-14406) to CL, and by NCI and NIEHS Grants UO1ES012771 and R21-ES15894 to JR.

Conflicts of interest statement

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Laboratory of Hormonal Carcinogenesis, Institute of Experimental Biology and Medicine (IBYME), National Research Council of Argentina (CONICET), Vuelta de Obligado 2490, C1428ADN, Buenos Aires, Argentina

    Victoria Wargon, Sebastián Giulianelli & Claudia Lanari

  2. Breast Cancer Laboratory, Fox Chase Cancer Center, Philadelphia, PA, USA

    Sandra V. Fernandez & Jose Russo

  3. Laboratorios Beta, IBYME, Buenos Aires, Argentina

    Mercedes Goin

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  1. Victoria Wargon
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  2. Sandra V. Fernandez
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  3. Mercedes Goin
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  4. Sebastián Giulianelli
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  5. Jose Russo
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  6. Claudia Lanari
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Correspondence to Claudia Lanari.

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Wargon, V., Fernandez, S.V., Goin, M. et al. Hypermethylation of the progesterone receptor A in constitutive antiprogestin-resistant mouse mammary carcinomas. Breast Cancer Res Treat 126, 319–332 (2011). https://doi.org/10.1007/s10549-010-0908-x

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