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Smad2 and Smad3 have opposing roles in breast cancer bone metastasis by differentially affecting tumor angiogenesis

Oncogene volume 29, pages 1351–1361 (2010)Cite this article

Abstract

Transforming growth factor (TGF)-β can suppress and promote breast cancer progression. How TGF-β elicits these dichotomous functions and which roles the principle intracellular effector proteins Smad2 and Smad3 have therein, is unclear. Here, we investigated the specific functions of Smad2 and Smad3 in TGF-β-induced responses in breast cancer cells in vitro and in a mouse model for breast cancer metastasis. We stably knocked down Smad2 or Smad3 expression in MDA-MB-231 breast cancer cells. The TGF-β-induced Smad3-mediated transcriptional response was mitigated and enhanced by Smad3 and Smad2 knockdown, respectively. This response was also seen for TGF-β-induced vascular endothelial growth factor (VEGF) expression. TGF-β induction of key target genes involved in bone metastasis, were found to be dependent on Smad3 but not Smad2. Strikingly, whereas knockdown of Smad3 in MDA-MB-231 resulted in prolonged latency and delayed growth of bone metastasis, Smad2 knockdown resulted in a more aggressive phenotype compared with control MDA-MB-231 cells. Consistent with differential effects of Smad knockdown on TGF-β-induced VEGF expression, these opposing effects of Smad2 versus Smad3 could be directly correlated with divergence in the regulation of tumor angiogenesis in vivo. Thus, Smad2 and Smad3 differentially affect breast cancer bone metastasis formation in vivo.

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Abbreviations

ALK:

activin receptor-like kinase

BLI:

bioluminescent imaging

BMP:

bone morphogenetic protein

CTGF:

connective tissue growth factor

EMT:

epithelial to mesenchymal transition

GFP:

green fluorescent protein

HIF-1α:

hypoxia inducible factor-1α

IL-11:

interleukin 11

miR RNAi:

micro RNA interference

MH:

Mad homology

N-T control:

non-targeting control

PAI-1:

plasminogen activator inhibitor-1

PlGF:

placenta growth factor

PTHrP:

parathyroid hormone-related protein

P-Smad:

phosphorylated Smad

R-Smad:

receptor-regulated Smad

Smad:

small phenotype and mothers against decapentaplegic related protein

TGF-β:

transforming growth factor-β

TβRII:

TGF-β type II receptor

VEGF:

vascular endothelial growth factor

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Acknowledgements

We thank Martine Deckers and Maarten van Dinther for their initial studies and members of our research group for help and suggestions during the course of this work. We are grateful to Lukas Hawinkels and Hein Verspaget for assistance with VEGF ELISA measurements, Kuber Sampath (Genzyme) for BMP6, Ken Iwata (OSI Pharmaceuticals) for TGF-β3 and Dr Philippe Clézardin for the MDA-MB-231 subclone BO2. This work was supported by the Dutch Cancer Society (UL 2005–3371), Ludwig Institute for Cancer Research and grants from the Sixth European Union Framework Programme, that is, the EpiplastCarcinoma Marie Curie RTN (project 005428), Angiotargeting (project 504743), BRECOSM (project 503224) and Tumor-Host Genomics (project 518198).

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Authors and Affiliations

  1. Department of Molecular Cell Biology and Centre for Biomedical Genetics, Leiden, The Netherlands

    M Petersen, E Pardali & P ten Dijke

  2. Department of Urology, Leiden University Medical Center, Leiden, The Netherlands

    M Petersen, G van der Horst, H Cheung, C van den Hoogen & G van der Pluijm

  3. Department of Endocrinology, Leiden University Medical Center, Leiden, The Netherlands

    G van der Pluijm

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  1. M Petersen
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Correspondence to G van der Pluijm.

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

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Petersen, M., Pardali, E., van der Horst, G. et al. Smad2 and Smad3 have opposing roles in breast cancer bone metastasis by differentially affecting tumor angiogenesis. Oncogene 29, 1351–1361 (2010). https://doi.org/10.1038/onc.2009.426

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