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Targeting MMP-9, uPAR, and cathepsin B inhibits invasion, migration and activates apoptosis in prostate cancer cells

Cancer Gene Therapy volume 17, pages 599–613 (2010)Cite this article

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Abstract

Prostate cancer is one of the most commonly diagnosed cancers and the second leading cause of cancer deaths in Americans. The high mortality rate is mainly attributed to the invasiveness and metastasis of advanced prostate cancer. Targeting the molecules involved in metastasis could be an effective mode of treatment for prostate cancer. In this study, the therapeutic potential of siRNA-mediated targeting of matrix metalloproteinase-9 (MMP-9), urokinase plasminogen activator receptor (uPAR), and cathepsin B (CB) in prostate cancer was carried out using single and bi-cistronic siRNA-expressing constructs. Downregulation of MMP-9, uPAR, and CB inhibited matrigel invasion, in vitro angiogenesis and wound-healing migration ability of PC3 and DU145 prostate cancer cell lines. In addition, the siRNA treatments induced apoptosis in the tumor cells as determined by TUNEL and DNA laddering assays. An attempt to elucidate the apoptotic pathway showed the involvement of FAS-mediated activation of caspases-8 and -7. Further, mice with orthotopic prostate tumors treated with siRNA-expressing vectors showed significant inhibition in tumor growth and migration. In conclusion, we report that the siRNA-mediated knockdown of MMP-9, uPAR, and CB inhibits invasiveness and migration of prostate cancer cells and leads to apoptosis both in vitro and in vivo.

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Abbreviations

CB:

cathepsin B

CMV:

cytomegalovirus

ECM:

extracellular matrix

FADD:

Fas-associated death domain

HMEC:

human microvascular endothelial cells

MMP:

matrix metalloproteases

PBS:

phosphate-buffered saline

RT-PCR:

reverse transcription polymerase chain reaction

SDS-PAGE:

sodium dodecyl sulphate-polyacrlyamide gel electrophoresis

uPAR:

urokinase-type plasminogen activator receptor

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Acknowledgements

We thank Shellee Abraham for manuscript preparation. We also thank Diana Meister and Sushma Jasti for manuscript review. This research was supported by National Cancer Institute Grant CA75557, CA116708, CA138409 and Caterpillar Inc., OSF St Francis Inc. Peoria, IL (to JSR). The contents of this manuscript are solely the responsibility of the authors and do not necessarily represent the official views of NIH.

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

    A K Nalla, B Gorantla, C S Gondi, S S Lakka & J S Rao

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

    J S Rao

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Correspondence to J S Rao.

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Dr Rao's work has been funded by NIH. The other authors declare no conflict of interest.

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Nalla, A., Gorantla, B., Gondi, C. et al. Targeting MMP-9, uPAR, and cathepsin B inhibits invasion, migration and activates apoptosis in prostate cancer cells. Cancer Gene Ther 17, 599–613 (2010). https://doi.org/10.1038/cgt.2010.16

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