The Ras-related protein, Rap1A, mediates thrombin-stimulated, integrin-dependent glioblastoma cell proliferation and tumor growth

Jacqueline Sayyah, Alena Bartakova, Nekeisha Nogal, Lawrence Quilliam, Dwayne G. Stupack, Joan Heller Brown

Research output: Contribution to journalArticle

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Abstract

Rap1 is a Ras family GTPase with a well documented role in ERK/MAP kinase signaling and integrin activation. Stimulation of the G-protein-coupled receptor PAR-1 with thrombin in human 1321N1 glioblastoma cells led to a robust increase in Rap1 activation. This response was sustained for up to 6 h and mediated through RhoA and phospholipase D (PLD). Thrombin treatment also induced a 5-fold increase in cell adhesion to fibronectin, which was blocked by down-regulating PLD or Rap1A or by treatment with a β1 integrin neutralizing antibody. In addition, thrombin treatment led to increases in phospho-focal adhesion kinase (tyrosine 397), ERK1/2 phosphorylation and cell proliferation, which were significantly inhibited in cells treated with β1 integrin antibody or Rap1A siRNA. To assess the role of Rap1A in tumor formation in vivo, we compared growth of 1321N1 cells stably expressing control, Rap1A or Rap1B shRNA in a mouse xenograft model. Deletion of Rap1A, but not of Rap1B, reduced tumor mass by >70% relative to control. Similar observations were made with U373MG glioblastoma cells in which Rap1A was down-regulated. Collectively, these findings implicate a Rap1A/β1 integrin pathway, activated downstream of G-protein-coupled receptor stimulation and RhoA, in glioblastoma cell proliferation. Moreover, our data demonstrate a critical role for Rap1A in glioblastoma tumor growth in vivo.

Original languageEnglish
Pages (from-to)17689-17698
Number of pages10
JournalJournal of Biological Chemistry
Volume289
Issue number25
DOIs
StatePublished - Jun 20 2014

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rap1 GTP-Binding Proteins
ras Proteins
Cell proliferation
Glioblastoma
Integrins
Thrombin
Tumors
Cell Proliferation
Phospholipase D
Growth
G-Protein-Coupled Receptors
Small Interfering RNA
Neoplasms
Chemical activation
Focal Adhesion Protein-Tyrosine Kinases
Phosphorylation
GTP Phosphohydrolases
Cell adhesion
Extracellular Signal-Regulated MAP Kinases
Neutralizing Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

The Ras-related protein, Rap1A, mediates thrombin-stimulated, integrin-dependent glioblastoma cell proliferation and tumor growth. / Sayyah, Jacqueline; Bartakova, Alena; Nogal, Nekeisha; Quilliam, Lawrence; Stupack, Dwayne G.; Brown, Joan Heller.

In: Journal of Biological Chemistry, Vol. 289, No. 25, 20.06.2014, p. 17689-17698.

Research output: Contribution to journalArticle

Sayyah, Jacqueline ; Bartakova, Alena ; Nogal, Nekeisha ; Quilliam, Lawrence ; Stupack, Dwayne G. ; Brown, Joan Heller. / The Ras-related protein, Rap1A, mediates thrombin-stimulated, integrin-dependent glioblastoma cell proliferation and tumor growth. In: Journal of Biological Chemistry. 2014 ; Vol. 289, No. 25. pp. 17689-17698.
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