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Ha-Ras transformation of MCF10A cells leads to repression of Singleminded-2s through NOTCH and C/EBPβ

Oncogene volume 28, pages 1561–1568 (2009)Cite this article

Abstract

We have previously shown that Singleminded-2s (SIM2s), a member of the basic helix-loop-helix Per-Arnt-Sim (bHLH/PAS) family of transcription factors, is downregulated in breast cancer samples and has tumor suppressor activity. However, the mechanism by which SIM2s is repressed in breast cancer cells has not been determined. In this study, we show that transformation of MCF10A cells by Harvey-Ras (Ha-Ras) induces CCAAT/enhance binding protein β (C/EBPβ) and activates the NOTCH signaling pathway to block SIM2s gene expression. NOTCH-mediated repression acts through a C-repeat binding factor 1 (CBF1)-independent mechanism, as introduction of CBF1 had no effect on SIM2s expression. Consistent with C/ebpβ-dependent inhibition of SIM2s, C/ebpβ−/− mouse mammary glands express high levels of SIM2s and reestablishment of C/ebpβ isoforms decreased SIM2s mRNA levels in C/ebpβ immortalized mammary epithelial cell lines. These studies illustrate a novel pathway of tumor suppressor gene silencing in Ha-Ras-transformed breast epithelial cells and identify SIM2s as a target of C/EBPβ and NOTCH signaling.

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Acknowledgements

We thank Bryan and Alana Welm (Huntsman Cancer Institute, Salt Lake City, UT, USA) for the Ha-Ras parent construct, Jeff Rosen (Baylor College of Medicine, Houston, TX, USA) for providing the C/ebpβ−/− mammary glands and Michael Lewis (Baylor College of Medicine, Houston, TX, USA) for critical reading of the paper. We also thank Keelan Anderson for technical assistance. This work was supported by awards from the National Cancer Institute RO1CA111551 (WWP), R01CA113795 (CAZ) and Howard Hughes Medical Institute Predoctoral Fellowship (TG).

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Authors and Affiliations

  1. Department of Integrative Biosciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA

    T L Gustafson, E Wellberg, B Laffin, L Schilling, R P Metz & W W Porter

  2. Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, MD, USA

    C A Zahnow

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  1. T L Gustafson
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  2. E Wellberg
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  3. B Laffin
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  4. L Schilling
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  7. W W Porter
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Correspondence to W W Porter.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Gustafson, T., Wellberg, E., Laffin, B. et al. Ha-Ras transformation of MCF10A cells leads to repression of Singleminded-2s through NOTCH and C/EBPβ. Oncogene 28, 1561–1568 (2009). https://doi.org/10.1038/onc.2008.497

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