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Wilms' tumor protein 1: an early target of progestin regulation in T-47D breast cancer cells that modulates proliferation and differentiation

Oncogene volume 27, pages 126–138 (2008)Cite this article

Abstract

Progesterone regulates the proliferation and differentiation of normal mammary epithelium. In breast cancer cells, progesterone and its synthetic analogs, progestins, induce long-term growth inhibition and differentiation. However, the mechanisms responsible are not fully understood. When T-47D breast cancer cells were treated with the synthetic progestin ORG 2058 (16α-ethoxy-21-hydroxy-19-norpregn-4-en-3,20-dione), all isoforms of Wilms' tumor protein 1 (Wt1) mRNA and protein were rapidly downregulated. We reasoned that the decrease in Wt1 levels may contribute to the long-term antiproliferative and differentiative effects of progestins as proliferation and differentiation are known end points of Wt1 action. Consistent with this idea, Wt1 small interfering RNA led to a decrease in S phase and cyclin D1 levels, and increased Oil-Red-O staining, indicating increased lipogenesis. Conversely, overexpression of Wt1 attenuated the decrease in S phase induced by ORG 2058 at 48–96 h. This was accompanied by the sustained expression of cyclin D1 despite progestin treatment, and increased levels of retinoblastoma (Rb) phosphorylation at sites targeted by cyclin D1-Cdk4 (Ser249/Thr252). Wt1 overexpression also attenuated the ORG 2058-mediated increase in fatty acid synthase levels and reduced lipogenesis. Thus, Wt1 downregulation was sufficient to mimic the effects of progestin and was necessary for complete progestin-mediated proliferative arrest and subsequent differentiation. Furthermore, Wt1 overexpression modulated the effects of progestins but not anti-estrogens or androgens. These results indicate that Wt1 is an important early target of progestins that regulates both proliferation and differentiation in breast cancer cells.

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Acknowledgements

We thank Mia Åkerfeldt, Ann Cornish, Lisa-Jane Hunter and Dr Alex Swarbrick for their contributions to some early experiments, and Gillian Lehrbach for providing the cell line RNA and protein. This research was supported by The National Health and Medical Research Council of Australia, the Cancer Institute NSW, the Australian Cancer Research Foundation (ACRF Unit for the Molecular Genetics of Cancer), the RT Hall Trust and The Cancer Council, NSW. CEC is a Cancer Institute NSW Scholar and EAM is a Cancer Institute NSW Fellow.

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  1. Cancer Research Program, Garvan Institute of Medical Research, Sydney, NSW, Australia

    C E Caldon, C S L Lee, R L Sutherland & E A Musgrove

  2. St Vincent's Clinical School, Faculty of Medicine, University of NSW, NSW, Australia

    C E Caldon, R L Sutherland & E A Musgrove

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  1. C E Caldon
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  2. C S L Lee
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Correspondence to E A Musgrove.

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Caldon, C., Lee, C., Sutherland, R. et al. Wilms' tumor protein 1: an early target of progestin regulation in T-47D breast cancer cells that modulates proliferation and differentiation. Oncogene 27, 126–138 (2008). https://doi.org/10.1038/sj.onc.1210622

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