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Calpain 2 is Required for Glioblastoma Cell Invasion: Regulation of Matrix Metalloproteinase 2

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Abstract

Invasion of glioblastoma cells significantly reduces the effectiveness of current treatments, highlighting the importance of understanding dispersal mechanisms and characteristics of the invasive population. Induction of calcium fluxes into glioblastoma cells by autocrine glutamate is critical for invasion. However, the target(s) by which calcium acts to stimulate the dispersal of glioblastoma cells is not clear. In this study, we tested the hypothesis that the calcium-activated protease calpain 2 is required for glioblastoma cell invasion. Knockdown of calpain 2 expression using shRNA or chemical inhibition of calpain activity reduced glioblastoma cell invasion by 90%. Interestingly, decreased expression of calpain 2 did not influence morphology or migration, suggesting regulation of invasion specific mechanisms. Consistent with this idea, 39% less extracellular MMP2 was measured from knockdown cells identifying one mechanism by which calpain 2 mediates glioblastoma cell invasion. This is the first report demonstrating that calpain 2 is required for glioblastoma cell invasion.

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Abbreviations

AMPA:

α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate

CL:

Collagen type IV

DMEM:

Dulbecco’s modified Eagle media

FBS:

Fetal bovine serum

FN:

Fibronectin

LN:

Laminin

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Acknowledgments

This work was supported by grants to J.A.G. from the Medical Research Foundation of Oregon, the General Research Fund of Oregon State University, and GM 63711 from the National Institute of General Medical Sciences, National Institutes of Health. This publication was made possible in part by the Cell Imaging and Analysis Facility and Services Core of the Environmental Health Sciences Center at Oregon State University from grant P30 ES00210, National Institute of Environmental Health Sciences, National Institutes of Health. Special thanks to Tamara S. Fraley for expert assistance with tissue culture and microscopy.

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  1. Department of Biochemistry and Biophysics, Oregon State University, ALS 2011, Corvallis, OR, 97331, USA

    Hyo Sang Jang, Sangeet Lal & Jeffrey A. Greenwood

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  1. Hyo Sang Jang
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  2. Sangeet Lal
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  3. Jeffrey A. Greenwood
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Correspondence to Jeffrey A. Greenwood.

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Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Graduate School of Oregon State University.

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Jang, H.S., Lal, S. & Greenwood, J.A. Calpain 2 is Required for Glioblastoma Cell Invasion: Regulation of Matrix Metalloproteinase 2. Neurochem Res 35, 1796–1804 (2010). https://doi.org/10.1007/s11064-010-0246-8

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