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RASSF1A inhibits estrogen receptor alpha expression and estrogen-independent signalling: implications for breast cancer development

Oncogene volume 31, pages 4912–4922 (2012)Cite this article

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Abstract

The Ras association domain family 1 isoform A (RASSF1A) is a tumor suppressor whose inactivation is implicated in the development of many human cancers, including breast carcinomas. Little is known about the tumor-suppressive function of RASSF1A in breast tissue and whether its inactivation is mechanistically involved in the initiation and progression of breast tumors. Here, we show that RASSF1A inhibits breast cancer growth in vivo, and suppresses estrogen receptor (ERα) expression and function. Reconstitution of RASSF1A in MCF7 cells led to decreased ERα levels and reduced sensitivity to estrogen (E2). Concomitantly, we observed decreased expression of Id1 as well as the E2-responsive genes Bcl-2 and c-Myc that cooperatively contribute to the immortalization and transformation of breast epithelial cells. This downregulation was associated with induction of cell-cycle arrest and senescence that constitute early barriers to cancer initiation and progression. Knockdown of ERα showed that downregulation of ERα suffices to increase senescence and inhibit expression of Bcl-2, c-Myc and Id1. However, enforced expression of ERα only partially rescued RASSF1A-mediated growth inhibition and senescence, suggesting that suppression of ERα expression and activity is not the only mechanism by which RASSF1A inhibits growth and survival of breast cancer cells. Ectopic expression of Bcl-2, c-Myc and Id1 had little or no effect on RASSF1A-mediated growth arrest, indicating that RASSF1A acts dominantly over these oncogenes. Mechanistically, RASSF1A was found to suppress ERα expression through Akt1. It also transiently inhibited ERα-induced Ras-MAPK activity after exposure of cells to E2. Together, our data show that RASSF1A acts as a tumor suppressor in ERα+ mammary epithelial cells, in part through inhibiting ERα expression and activity. These findings suggest that RASSF1A has a key role in suppressing the transformation of human breast epithelial cells and ERα+ breast cancer initiation.

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Acknowledgements

We are grateful to Selma Huber and workers in the KIT rodent facility and Gitta Flaig for excellent technical assistance. We thank Robert Weinberg for kindly providing pBabe-neo; William Hahn for providing pBabe-Neo-Myr-Flag-AKT1 and pWZL Blast myc and Jean Zhao for pWZL Neo Myr Flag AKT3. pMIG Bcl-2 was created in the laboratory of Stanley Korsmeyer and was kindly provided from Dana Farber Cancer Institute. This study was supported by Deutsche Forschungsgemeinschaft grant TH1523/1-2 to ST and MAIFOR (ST and MS).

Author contributions: ST designed research; ST, AS and MS performed research, MS and AS contributed new reagents/analytic tools, ST analyzed data; ST and JPS wrote the paper.

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Authors and Affiliations

  1. Centre for Biomedicine and Medical Technology Mannheim (CBTM), Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany

    S Thaler & J P Sleeman

  2. Department of Obestrics and Gynecology, Johannes Gutenberg University, Mainz, Germany

    M Schmidt

  3. Department of Pathology, Johannes Gutenberg University, Mainz, Germany

    A Schad

  4. KIT Campus Nord, Institut für Toxikologie und Genetik, Karlsruhe, Germany

    J P Sleeman

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Correspondence to S Thaler.

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Thaler, S., Schmidt, M., Schad, A. et al. RASSF1A inhibits estrogen receptor alpha expression and estrogen-independent signalling: implications for breast cancer development. Oncogene 31, 4912–4922 (2012). https://doi.org/10.1038/onc.2011.658

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