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Cancer Chemotherapy and Pharmacology

, Volume 56, Issue 6, pp 585–593 | Cite as

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

  • Keisuke Yamaguchi
  • Mark D. Richardson
  • Darrell D. Bigner
  • Madan M. KwatraEmail author
Original Article

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.

Keywords

Substance P receptor Epidermal growth factor receptor (EGFR) Src ERK1/2 Protein kinase C (PKC) Glioblastoma 

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

Notes

Acknowledgements

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

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Keisuke Yamaguchi
    • 1
  • Mark D. Richardson
    • 2
  • Darrell D. Bigner
    • 3
    • 4
  • Madan M. Kwatra
    • 2
    • 4
    • 5
    • 6
    • 2
    Email author
  1. 1.Department of Anesthesiology and Pain MedicineJuntendo University School of MedicineTokyoJapan
  2. 2.Department of AnesthesiologyDuke University Medical CenterDurhamUSA
  3. 3.Department of PathologyDuke University Medical CenterDurhamUSA
  4. 4.Duke Comprehensive Cancer CenterDuke University Medical CenterDurhamUSA
  5. 5.Department of Pharmacology and Cancer BiologyDuke University Medical CenterDurhamUSA
  6. 6.Center for the Study of Aging and Human DevelopmentDuke University Medical CenterDurhamUSA

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