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Signal transduction through substance P receptor in human glioblastoma cells: roles for Src and PKCδ

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Abstract

Substance P receptor (SPR), a G protein-coupled receptor (GPCR), is found in human glioblastomas, and has been implicated in their growth. Consistent with a role for SPR in cell growth, activation of SPR in U373 MG human glioblastoma cells leads to the phosphorylation of mitogen-activated protein kinases [extracellular signal-regulated kinase 1 and 2 (ERK1/2)] and stimulation of cell proliferation. The purpose of the present study was to elucidate the pathway through which these actions occur. Using either the epidermal growth factor receptor (EGFR) kinase inhibitor, AG 1478, or a small-interfering RNA (siRNA) directed against human EGFR, we found that transactivation of EGFR by SPR is only marginally involved in SP-dependent ERK1/2 phosphorylation. Src, however, is shown to be a major component of SPR signaling because the Src kinase inhibitor, PP2, and a kinase-dead Src mutant both inhibit SP-dependent ERK1/2 phosphorylation. We also report that SPR stimulates the phosphorylation of protein kinase Cδ(PKCδ), and that this stimulation is blocked by PP2. SP-dependent ERK1/2 phosphorylation is also blocked by rottlerin, a PKCδ inhibitor, and the calcium scavenger, BAPTA/AM. Finally, rottlerin and PP2 were both found to inhibit the growth of several glioblastoma cell lines, underscoring the potential of these agents to block glioblastoma growth.

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Abbreviations

SPR:

Substance P receptor

GPCR:

G protein-coupled receptor

SP:

Substance P

PKC:

Protein kinase C

ERK1/2:

Extracellular signal-regulated kinase 1 and 2

EGFR:

Epidermal growth factor receptor

BAPTA/AM:

1,2-bis-(o-Aminophenoxy)ethane-N, N, N′, N′-tetraacetic acid tetra-(acetoxymethyl) ester

PP2:

4-Amino-5-(4-chlorphenyl)-7-(tbutyl) pyrazolo[3,4-d]pyrimidine

PP3:

4-Amino-7-phenylpyrazol[3,4-d]pyrimidine

EGF:

Epidermal growth factor

PMA:

Phorbol-12-myristate-13-acetate

GF109203X:

Bisindoylmaleimide I

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Acknowledgements

This work was supported in part by National Institutes of Health Grant NS 33405 (to M.M.K.).

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Authors and Affiliations

  1. Department of Anesthesiology and Pain Medicine, Juntendo University School of Medicine, Tokyo, Japan

    Keisuke Yamaguchi

  2. Department of Anesthesiology, Duke University Medical Center, 3094, Durham, NC, 22710, USA

    Mark D. Richardson, Madan M. Kwatra & Madan M. Kwatra

  3. Department of Pathology, Duke University Medical Center, Durham, NC, 27710, USA

    Darrell D. Bigner

  4. Duke Comprehensive Cancer Center, Duke University Medical Center, Durham, NC, 27710, USA

    Darrell D. Bigner & Madan M. Kwatra

  5. Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, 27710, USA

    Madan M. Kwatra

  6. Center for the Study of Aging and Human Development, Duke University Medical Center, Durham, NC, 27710, USA

    Madan M. Kwatra

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  1. Keisuke Yamaguchi
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Correspondence to Madan M. Kwatra.

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Yamaguchi, K., Richardson, M.D., Bigner, D.D. et al. Signal transduction through substance P receptor in human glioblastoma cells: roles for Src and PKCδ. Cancer Chemother Pharmacol 56, 585–593 (2005). https://doi.org/10.1007/s00280-005-1030-3

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  • DOI: https://doi.org/10.1007/s00280-005-1030-3

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