Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling

Nirmala Chandrasekar, Sanjeeva Mohanam, S. Sajani Lakka, Dzung H. Dinh, William C. Olivero, Meena Gujrati, Jasti S. Rao

Research output: Contribution to journalArticle

Abstract

Purpose: Tumor vasculature provides the infrastructure by which malignant tissue can be nourished; therefore, targeting angiogenesis may be an effective means of treating cancer. We showed previously that SNB19 glioblastoma cells modulate bovine retinal endothelial cells in cocultures to form capillary-like network structures, that matrix metalloproteinase-9 (MMP-9) expression is critical for endothelial morphogenesis, and that MMP-9 expression in glioblastoma cells is regulated by extracellular signal-regulated kinase-1 (ERK-1). In the present study, we investigated whether interfering with the activation of this mitogen-activated protein (MAP) kinase would repress MMP-9 synthesis and inhibit capillary formation. Experimental Design: Cocultures of bovine retinal endothelial and SNB19 cells were analyzed for MMP-9 secretion, and phospho- and total ERK levels. These cocultures were treated with PD98059, a specific inhibitor of MAP/ERK kinase 1, or transfected with dominant-negative ERK-1 mutant containing expression vector. Alterations in capillary-like structure formation, and actin cytoskeleton and secretion of vascular endothelial growth factor (VEGF), MMP-9, and tissue inhibitor of metalloproteinase-1 were determined by immunofluorescence, gelatin zymography, and Western blotting. Results: We found that inhibition of the ERK-1/2 pathway with PD98059 abrogated glial cell-mediated capillary formation by the endothelial cells and reduced the levels of MMP-9 in the coculture. Strikingly, the abrogation of MAP kinase signaling by a dominant-negative ERK-1 mutant inhibited glial-induced capillary network formation by reducing VEGF levels and MMP-9 activity and increasing the levels of tissue inhibitor of metalloproteinase-1. Inhibition of ERK activity also disrupted the formation of the actin cytoskeleton, a prerequisite for endothelial cell migration. Conclusion: The mechanism underlying activation of ERK is involved in reorganization of the actin cytoskeleton, and induction of VEGF and MMP-9, thereby stimulating endothelial cell morphogenesis. These studies clearly provide experimental evidence that ERK inhibition diminishes glial-induced endothelial-cell morphogenesis; therefore, interfering with ERK signaling may be a viable approach to target angiogenesis.

Original languageEnglish (US)
Pages (from-to)2342-2349
Number of pages8
JournalClinical Cancer Research
Volume9
Issue number6
StatePublished - Jun 1 2003

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Extracellular Signal-Regulated MAP Kinases
Matrix Metalloproteinase 9
Morphogenesis
Neuroglia
Mitogen-Activated Protein Kinase 3
Endothelial Cells
Coculture Techniques
Mitogen-Activated Protein Kinases
Actin Cytoskeleton
Vascular Endothelial Growth Factor A
Tissue Inhibitor of Metalloproteinase-1
Glioblastoma
Matrix Metalloproteinase Inhibitors
Mitogen-Activated Protein Kinase 1
Gelatin
Cell Movement
Fluorescent Antibody Technique
Neoplasms
Research Design
Western Blotting

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Chandrasekar, N., Mohanam, S., Lakka, S. S., Dinh, D. H., Olivero, W. C., Gujrati, M., & Rao, J. S. (2003). Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling. Clinical Cancer Research, 9(6), 2342-2349.

Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling. / Chandrasekar, Nirmala; Mohanam, Sanjeeva; Lakka, S. Sajani; Dinh, Dzung H.; Olivero, William C.; Gujrati, Meena; Rao, Jasti S.

In: Clinical Cancer Research, Vol. 9, No. 6, 01.06.2003, p. 2342-2349.

Research output: Contribution to journalArticle

Chandrasekar, N, Mohanam, S, Lakka, SS, Dinh, DH, Olivero, WC, Gujrati, M & Rao, JS 2003, 'Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling', Clinical Cancer Research, vol. 9, no. 6, pp. 2342-2349.
Chandrasekar N, Mohanam S, Lakka SS, Dinh DH, Olivero WC, Gujrati M et al. Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling. Clinical Cancer Research. 2003 Jun 1;9(6):2342-2349.
Chandrasekar, Nirmala ; Mohanam, Sanjeeva ; Lakka, S. Sajani ; Dinh, Dzung H. ; Olivero, William C. ; Gujrati, Meena ; Rao, Jasti S. / Glial cell-induced endothelial morphogenesis is inhibited by interfering with extracellular signal-regulated kinase signaling. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 6. pp. 2342-2349.
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abstract = "Purpose: Tumor vasculature provides the infrastructure by which malignant tissue can be nourished; therefore, targeting angiogenesis may be an effective means of treating cancer. We showed previously that SNB19 glioblastoma cells modulate bovine retinal endothelial cells in cocultures to form capillary-like network structures, that matrix metalloproteinase-9 (MMP-9) expression is critical for endothelial morphogenesis, and that MMP-9 expression in glioblastoma cells is regulated by extracellular signal-regulated kinase-1 (ERK-1). In the present study, we investigated whether interfering with the activation of this mitogen-activated protein (MAP) kinase would repress MMP-9 synthesis and inhibit capillary formation. Experimental Design: Cocultures of bovine retinal endothelial and SNB19 cells were analyzed for MMP-9 secretion, and phospho- and total ERK levels. These cocultures were treated with PD98059, a specific inhibitor of MAP/ERK kinase 1, or transfected with dominant-negative ERK-1 mutant containing expression vector. Alterations in capillary-like structure formation, and actin cytoskeleton and secretion of vascular endothelial growth factor (VEGF), MMP-9, and tissue inhibitor of metalloproteinase-1 were determined by immunofluorescence, gelatin zymography, and Western blotting. Results: We found that inhibition of the ERK-1/2 pathway with PD98059 abrogated glial cell-mediated capillary formation by the endothelial cells and reduced the levels of MMP-9 in the coculture. Strikingly, the abrogation of MAP kinase signaling by a dominant-negative ERK-1 mutant inhibited glial-induced capillary network formation by reducing VEGF levels and MMP-9 activity and increasing the levels of tissue inhibitor of metalloproteinase-1. Inhibition of ERK activity also disrupted the formation of the actin cytoskeleton, a prerequisite for endothelial cell migration. Conclusion: The mechanism underlying activation of ERK is involved in reorganization of the actin cytoskeleton, and induction of VEGF and MMP-9, thereby stimulating endothelial cell morphogenesis. These studies clearly provide experimental evidence that ERK inhibition diminishes glial-induced endothelial-cell morphogenesis; therefore, interfering with ERK signaling may be a viable approach to target angiogenesis.",
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AU - Olivero, William C.

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AU - Rao, Jasti S.

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N2 - Purpose: Tumor vasculature provides the infrastructure by which malignant tissue can be nourished; therefore, targeting angiogenesis may be an effective means of treating cancer. We showed previously that SNB19 glioblastoma cells modulate bovine retinal endothelial cells in cocultures to form capillary-like network structures, that matrix metalloproteinase-9 (MMP-9) expression is critical for endothelial morphogenesis, and that MMP-9 expression in glioblastoma cells is regulated by extracellular signal-regulated kinase-1 (ERK-1). In the present study, we investigated whether interfering with the activation of this mitogen-activated protein (MAP) kinase would repress MMP-9 synthesis and inhibit capillary formation. Experimental Design: Cocultures of bovine retinal endothelial and SNB19 cells were analyzed for MMP-9 secretion, and phospho- and total ERK levels. These cocultures were treated with PD98059, a specific inhibitor of MAP/ERK kinase 1, or transfected with dominant-negative ERK-1 mutant containing expression vector. Alterations in capillary-like structure formation, and actin cytoskeleton and secretion of vascular endothelial growth factor (VEGF), MMP-9, and tissue inhibitor of metalloproteinase-1 were determined by immunofluorescence, gelatin zymography, and Western blotting. Results: We found that inhibition of the ERK-1/2 pathway with PD98059 abrogated glial cell-mediated capillary formation by the endothelial cells and reduced the levels of MMP-9 in the coculture. Strikingly, the abrogation of MAP kinase signaling by a dominant-negative ERK-1 mutant inhibited glial-induced capillary network formation by reducing VEGF levels and MMP-9 activity and increasing the levels of tissue inhibitor of metalloproteinase-1. Inhibition of ERK activity also disrupted the formation of the actin cytoskeleton, a prerequisite for endothelial cell migration. Conclusion: The mechanism underlying activation of ERK is involved in reorganization of the actin cytoskeleton, and induction of VEGF and MMP-9, thereby stimulating endothelial cell morphogenesis. These studies clearly provide experimental evidence that ERK inhibition diminishes glial-induced endothelial-cell morphogenesis; therefore, interfering with ERK signaling may be a viable approach to target angiogenesis.

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