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Anthrax toxin receptor 2 is expressed in murine and tumor vasculature and functions in endothelial proliferation and morphogenesis

Oncogene volume 29, pages 789–801 (2010)Cite this article

Abstract

The Capillary Morphogenesis Gene 2 (CMG2) gene encodes an Anthrax toxin receptor (ANTXR2), but the normal physiological function is not known. ANTXR2/CMG2 was originally identified as a result of up-regulation during capillary morphogenesis of endothelial cells (ECs) cultured in vitro. We explored the hypothesis that key steps of the angiogenic process are either dependent or are influenced by ANTXR2/CMG2 activity. We describe the expression pattern of ANTXR2/CMG2 in several murine tissues and in normal breast and breast tumors. Endothelial expression was found in all of the tissues analyzed, in cultured ECs and in breast tumor vessels; however, ANTXR2/CMG2 expression was not restricted to this cell type. To assess potential angiogenic function, we used RNA interference to achieve significant reduction of ANTXR2/CMG2 expression in cultured human umbilical venous endothelial cells (HUVECs). Reduced ANTXR2/CMG2 expression resulted in significant inhibition of proliferation and reduced capacity of ECs to form capillary-like networks in vitro, whereas overexpression of ANTXR2/CMG2 in HUVEC increased proliferation and capillary-like network formation. Little change in migration of ECs was observed on knockdown or overexpression. We conclude that ANTXR2/CMG2 functions to promote endothelial proliferation and morphogenesis during sprouting angiogenesis, consistent with the endothelial expression of ANTXR2/CMG2 in several vascular beds.

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Abbreviations

CMG2:

capillary morphogenesis gene 2

ANTXR2:

anthrax toxin receptor 2

HUVEC:

human umbilical venous endothelial cell

EC:

endothelial cell

VEGF:

vascular endothelial growth factor

EGF:

epidermal growth factor

ANTXR1:

anthrax toxin receptor 1

TEM8:

tumor endothelial marker 8

SFM:

serum-free medium

shRNA:

short hairpin RNA

PA:

protective antigen

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Acknowledgements

This work was supported by NIH RO1 HL62454 (JKK) and NIH RO1 AI064654 (JKK). CVR was supported by endocrinology training grant DK07328 and by a fellowship from the DOD Breast Cancer Program X81WH-06-1-0355. JY was supported by NIH grant AI48489. We thank Dr Yasuhiro Funahashi, Ayelet Spitzer, and Eunice Kim for technical support, Dr David Owens for the gift of the antibodies and critical reading of the paper, and Dr Darrell Yamashiro for help with preparation of the figures.

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Authors and Affiliations

  1. OB/GYN, Columbia University Medical Center, New York, NY, USA

    C V Reeves, J Dufraine & J Kitajewski

  2. Department of Pathology, Irving Cancer Research Center, Columbia University Medical Center, New York, NY, USA

    C V Reeves, J Dufraine & J Kitajewski

  3. Infectious Disease Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA

    J A T Young

  4. Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, New York, NY, USA

    J Kitajewski

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  1. C V Reeves
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  2. J Dufraine
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Correspondence to J Kitajewski.

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

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Reeves, C., Dufraine, J., Young, J. et al. Anthrax toxin receptor 2 is expressed in murine and tumor vasculature and functions in endothelial proliferation and morphogenesis. Oncogene 29, 789–801 (2010). https://doi.org/10.1038/onc.2009.383

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