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The LIM-only factor LMO4 regulates expression of the BMP7 gene through an HDAC2-dependent mechanism, and controls cell proliferation and apoptosis of mammary epithelial cells

Oncogene volume 26, pages 6431–6441 (2007)Cite this article

Abstract

The nuclear LIM-only protein 4 (LMO4) is upregulated in breast cancer, especially estrogen receptor-negative tumors, and its overexpression in mice leads to hyperplasia and tumor formation. Here, we show that deletion of LMO4 in the mammary glands of mice leads to impaired lobuloalveolar development due to decreased epithelial cell proliferation. With the goal of discovering potential LMO4-target genes, we also developed a conditional expression system in MCF-7 cells for both LMO4 and a dominant negative (DN) form of its co-regulator, cofactor of LIM domains (Clim/Ldb/Nli). We then used DNA microarrays to identify genes responsive to LMO4 and DN-Clim upregulation. One of the genes common to both data sets was bone morphogenic protein 7 (BMP7), whose expression is also significantly correlated with LMO4 transcript levels in a large dataset of human breast cancers, suggesting that BMP7 is a bona fide target gene of LMO4 in breast cancer. Inhibition of BMP7 partially blocks the effects of LMO4 on apoptosis, indicating that BMP7 mediates at least some functions of LMO4. Gene transfer studies show that LMO4 regulates the BMP7 promoter, and chromatin immunoprecipitation studies show that LMO4 and its cofactor Clim2 are recruited to the BMP7 promoter. Furthermore, we demonstrate that HDAC2 recruitment to the BMP7 promoter is inhibited by upregulation of LMO4 and that HDAC2 knockdown upregulates the promoter. These studies suggest a novel mechanism of action for LMO4: LMO4, Clim2 and HDAC2 are part of a transcriptional complex, and increased LMO4 levels can disrupt the complex, leading to decreased HDAC2 recruitment and increased promoter activity.

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Acknowledgements

This study was supported by NIH Grant AR44882 and the Irving Weinstein Foundation (BA); the Breast Cancer Research Program of the US Army Medical Research and Material Command (NW and ZL); and the California Breast Cancer Research Program (ZY and XX). We thank Hung Lin for PCR, Julie Fernandez for cell culture, UCI Microarray Core Facility for microarray experiments, and Rainer Brachman, Xing Dai, Steve Lipkin and Marian Waterman for reading the manuscript.

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Authors and Affiliations

  1. Departments of Medicine and Biological Chemistry, University of California, Irvine, CA, USA

    N Wang, K K Lin, Z Lu, K S Lam, R Newton, X Xu, Z Yu & B Andersen

  2. Department of Medicine, University of California, San Diego, La Jolla, CA, USA

    G N Gill

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  1. N Wang
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  2. K K Lin
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Correspondence to B Andersen.

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

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Wang, N., Lin, K., Lu, Z. et al. The LIM-only factor LMO4 regulates expression of the BMP7 gene through an HDAC2-dependent mechanism, and controls cell proliferation and apoptosis of mammary epithelial cells. Oncogene 26, 6431–6441 (2007). https://doi.org/10.1038/sj.onc.1210465

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