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Regulation of TGFβ1-mediated growth inhibition and apoptosis by RUNX2 isoforms in endothelial cells

Oncogene volume 23, pages 4722–4734 (2004)Cite this article

Abstract

Runx transcription factors regulate viral growth, hematopoiesis, bone formation, angiogenesis, and gastric epithelial development through specific DNA-binding motifs on target gene promoters. Vascular endothelial cells (ECs) express RUNX genes that are activated by angiogenic factors. The RUNX2 gene also activates the vascular endothelial growth factor promoter. Alternatively spliced forms of RUNX genes have been described, but their functions in angiogenesis have not been elucidated. In this study, expression of a novel alternatively spliced variant of RUNX2 (RUNX2Δ8), lacking the region encoded by exon 8, was detected in aortic tissue undergoing angiogenesis in vitro and in ECs. Expression of RUNX2 and RUNX2Δ8 increased in vascular sprouts concomitant with expression of cellular proteases and cytokines known to mediate angiogenesis. RUNX2 DNA-binding activity was expressed in proliferating but not quiescent ECs. Ectopic expression of RUNX2 in ECs increased cell sprouting, cell proliferation, DNA synthesis, and phosphorylation of phosphorylated retinoblastoma relative to control transfectants while RUNX2, but not RUNX2Δ8 transfectants, acquired resistance to growth inhibition by transforming growth factor (TGFβ1). Furthermore, RUNX2Δ8-transfected cells were more sensitive to TGFβ1-induced apoptosis than RUNX2 transfectants. Consistent with these data, the RUNX2 gene was a strong repressor of the promoter of the cyclin-dependent kinase inhibitor, p21CIP1, while RUNX2Δ8 was a competitive inhibitor of RUNX2 and exhibited weak repression activity. These results support the hypothesis that ECs regulate growth and apoptosis, in part, by alternative splicing events in the RUNX2 transcription factor to affect the TGFβ1 signaling pathway. The exon 8 domain of RUNX2 may contribute to the strong repression activity of RUNX2 for some target gene promoters.

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Abbreviations

EC:

endothelial cell

TGFβ1:

transforming growth factor-β

VEGF:

vascular endothelial growth factor

FGF:

fibroblast growth factor

IGF-1:

insulin-like growth factor-1

RHD:

Runt homology domain

uPA:

urokinase plasminogen activator

NF-κB:

nuclear factor-κB

CP:

cyclophilin

MT1-MMP:

membrane-type metalloproteinase-1

EMSA:

electrophoretic mobility-shift assay

BAEC:

bovine aortic endothelial cells

HBME:

human bone marrow endothelial cells. RUNX2 indicates the human gene whereas Runx2 indicates the mouse gene. In reference to general cases, Runx is used

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Acknowledgements

We thank Dr Yoshiaki Ito for providing the PEBP2aA (RUNX2) cDNA clone, Dr Bert Vogelstein for the p21CIP1-promoter vector, Mr. Jay Hunter for laboratory management, and Dr Jeffrey Hasday for his helpful comments in the preparation of the manuscript. This study was supported, in part, by an American Heart Association Grant-in-Aid (AHA 0151434U) and a National Institutes of Health grant (CA95350-01) to A.P. Michele Vitolo is the recipient of an American Heart Association Pre-doctoral Fellowship (AHA 0215217U).

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  1. Lixin Sun

    Present address: Neuro-Oncology Branch, National Cancer Institute, NIH, Bldg 36/3B02, Bethesda, MD, 20892, USA

Authors and Affiliations

  1. Greenebaum Cancer Center, University of Maryland School of Medicine, 655 W. Baltimore Street, Baltimore, 21201, MD, USA

    Lixin Sun, Ian E Anglin & Antonino Passaniti

  2. Department of Biochemistry & Molecular Biology, University of Maryland School of Medicine, Baltimore, 21201, MD, USA

    Michele I Vitolo & Antonino Passaniti

  3. Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, 21201, MD, USA

    Meng Qiao

  4. Department of Pathology, University of Maryland School of Medicine, Baltimore, 21201, MD, USA

    Ian E Anglin & Antonino Passaniti

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Correspondence to Antonino Passaniti.

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Sun, L., Vitolo, M., Qiao, M. et al. Regulation of TGFβ1-mediated growth inhibition and apoptosis by RUNX2 isoforms in endothelial cells. Oncogene 23, 4722–4734 (2004). https://doi.org/10.1038/sj.onc.1207589

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