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Cathepsin B and uPAR knockdown inhibits tumor-induced angiogenesis by modulating VEGF expression in glioma

Cancer Gene Therapy volume 18, pages 419–434 (2011)Cite this article

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Abstract

Angiogenesis, which is the process of sprouting of new blood vessels from pre-existing vessels, is vital for tumor progression. Proteolytic remodeling of extracellular matrix is a key event in vessel sprouting during angiogenesis. Urokinase type plasminogen activator receptor (uPAR) and cathepsin B are both known to be overexpressed and implicated in tumor angiogenesis. In the present study, we observed that knockdown of uPAR and cathepsin B using puPAR (pU), pCathepsin B (pC), and a bicistronic construct of uPAR and cathepsin B (pCU) caused significant inhibition of angiogenesis by disrupting the janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway-dependent expression of vascular endothelial growth factor (VEGF). Further, transcriptional suppression of uPAR and cathepsin B inhibited tumor-induced migration, proliferation of endothelial cells and decreased tumor-promoted expression of VEGF receptor-2, Rac1, gp91phox, cyclin D1, cyclin dependent kinase 4 and p-Rb in human dermal microvascular endothelial cell. Furthermore, U251 and SNB19 xenograft tissue sections from nude mice treated with pCU showed reduced expression of VEGF and CD31, which is a blood vessel visualization marker. Overall, results revealed that knockdown of uPAR and cathepsin B inhibited tumor-induced angiogenesis by disrupting the JAK/STAT pathway-dependent expression of VEGF. These data provide new insight in characterizing the pathways involved in the angiogenic cascade and for the identification of novel target proteins for use in therapeutic intervention for gliomas.

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Abbreviations

HMEC:

human dermal microvascular endothelial cell

JAK:

janus kinase

PBS:

phosphate-buffered saline

STAT:

signal transducer and activator of transcription

uPAR:

urokinase-type plasminogen activator receptor

VEGF:

vascular endothelial growth factor

NADPH:

nicotinamide adenine dinucleotide phosphate

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Acknowledgements

We thank Noorjehan Ali for technical assistance, Shellee Abraham for manuscript preparation and Diana Meister and Sushma Jasti for manuscript review. This research was supported by the National Cancer Institute Grant CA116708. The contents of this manuscript are solely the responsibility of the authors and do not necessarily represent the official views of NIH. Dr JS Rao's work has been funded by NIH.

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Authors and Affiliations

  1. Department of Cancer Biology and Pharmacology, University of Illinois College of Medicine, Peoria, IL, USA

    R R Malla, S Gopinath, C S Gondi, K Alapati & J S Rao

  2. Department of Neurosurgery, University of Illinois College of Medicine, Peoria, IL, USA

    D H Dinh & J S Rao

  3. Department of Pathology, University of Illinois College of Medicine, Peoria, IL, USA

    M Gujrati

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  1. R R Malla
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Malla, R., Gopinath, S., Gondi, C. et al. Cathepsin B and uPAR knockdown inhibits tumor-induced angiogenesis by modulating VEGF expression in glioma. Cancer Gene Ther 18, 419–434 (2011). https://doi.org/10.1038/cgt.2011.9

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