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Inhibition of Necl-5 (CD155/PVR) reduces glioblastoma dispersal and decreases MMP-2 expression and activity

  • Laboratory Investigation - Human/Animal Tissue
  • Published:
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Abstract

Patients afflicted with glioblastoma (GBM) have poor survival due to dispersive invasion throughout the brain. Necl-5, a cell surface receptor for vitronectin, is expressed in GBM but not normal brain. In several GBM cell lines Necl-5 promotes migration and invasion but the mechanism is poorly understood. In this study, we show that knockdown of Necl-5 by RNAi results in markedly decreased invasion of A172 GBM cells in a 3-dimensional matrix. There is a concomitant decrease in the expression and activity of matrix metalloproteinase-2 (MMP-2), a known factor in GBM invasion and disease severity. Knockdown of Necl-5 diminishes basal activation of Akt, an established mediator of MMP-2 expression in gliomas. Knockdown of Necl-5 also limits the maximal Akt activation in response to vitronectin, which requires the activity of Integrin-linked kinase (ILK). During migration, Necl-5, Akt and ILK co-localize at focal contacts at the leading edge of the plasma membrane, suggesting that these molecules may act to integrate Akt signaling at the leading edge to induce MMP-2 expression. By virtue of its restricted expression in GBM and its role in invasion, Necl-5 may be an attractive target for limiting MMP-2 production in glioblastoma, and therefore limiting dispersal.

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Acknowledgments

We thank Jean Stewart for technical assistance. We also thank Drs. Rob Jackson and Alenka Lovy-Wheeler of the Tufts Center for Neuroscience Research (P30 NS047243) for assistance with confocal imaging. The work was funded by grants from the Goldhirsh Foundation and the National Cancer Institute (CA116642).

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  1. Tufts University School of Medicine, Boston, MA, USA

    Brian M. Enloe & Daniel G. Jay

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  1. Brian M. Enloe
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Correspondence to Daniel G. Jay.

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11060_2010_323_MOESM1_ESM.ppt

Supplemental Fig. S1 HCT116 colon cancer cells were transfected with either control or Necl-5 siRNA (75 nM) using Oligofectamine and following the manufacturer’s directions. Spheroids were made using the hanging drop method as described by Del Duca et al. [21]. Spheroids were implanted into 1.75 mg/ml collagen I and photographed just after implantation (T0) and then 7 days later (T 7ds). Shown are representative micrographs of invasive control and non-invasive Necl-5 depleted spheroids. Nine of each spheroids were implanted, and 9/9 control spheroids were invasive, while 7/9 Necl-5 depleted spheroids were non-invasive. (PPT 22657 kb)

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Enloe, B.M., Jay, D.G. Inhibition of Necl-5 (CD155/PVR) reduces glioblastoma dispersal and decreases MMP-2 expression and activity. J Neurooncol 102, 225–235 (2011). https://doi.org/10.1007/s11060-010-0323-5

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  • DOI: https://doi.org/10.1007/s11060-010-0323-5

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