Vascular Endothelial Growth Factor (VEGF) increases matrix metalloproteinase production by human microvascular endothelial cells in vitro

W. J. Lamoreaux, M. E C Fitzgerald, Anton Reiner, Karen Hasty, S. T. Charles

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Abstract

Purpose: Proliferation and migration of vascular endothelial cells (EC) are characteristic of retinal neovascularization during proliferative diabetic retinopathy (PDR). Proliferation of EC has been shown to be induced by vascular endothelial growth factor (VEGF). To facilitate migration through the extracellular matrix surrounding these endothelial cells, matrix metalloproteinases (MMPs) may be produced. The MMPs are a family of enzymes that degrade the extracellular matrix and include the collagenases, gelatinases A and B (72 kDa and 92 kDa gelatinases, respectively) and the stromelysins. The MMPs must be activated by proteolytic cleavage in vivo or by organomercurials or sodium dodecyl sulfate (SDS) in vitro. We wished to determine if VEGF could alter in vitro MMP production by human microvascular EC. Methods: Commercial microvascular EC isolated from human lungs (hMVEC-L; Clonetics: San Diego) were grown to confluency. Cells were plated on plastic cell culture ware with 10 without 10 ng/ml VEGF. Media covering the cells were removed after 24 hr and aralyzed by zymography to determine MMP production. Results: The control (untreated) hMVEC-L produced a gelatinase with an apparent Mr of 63 kDa. This enzyme is similar in molecular weight to activated gelatinase A and may represent the result of SDS activation in vitro. Treatment of an equal number of hMVEC-L with 10 ng/ml VEGF resulted in an increase in gelatinolytic activity by at least 100%, indicating the presence of more gelatinase A. Conclusions: VEGF activation of microvascular endothelial cell proliferation is coordinated with an increase in gelatinase A production by these cells, indicating a possible mechanism for the coordination in proliferation and migration during PDR and other retinal pathologies that exhibit neovascularization.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996

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Matrix Metalloproteinases
Vascular Endothelial Growth Factor A
Endothelial Cells
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Diabetic Retinopathy
Sodium Dodecyl Sulfate
Extracellular Matrix
Retinal Neovascularization
Matrix Metalloproteinase 3
Gelatinases
Collagenases
Enzymes
Plastics
In Vitro Techniques
Cell Culture Techniques
Molecular Weight
Cell Proliferation
Pathology
Lung

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

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title = "Vascular Endothelial Growth Factor (VEGF) increases matrix metalloproteinase production by human microvascular endothelial cells in vitro",
abstract = "Purpose: Proliferation and migration of vascular endothelial cells (EC) are characteristic of retinal neovascularization during proliferative diabetic retinopathy (PDR). Proliferation of EC has been shown to be induced by vascular endothelial growth factor (VEGF). To facilitate migration through the extracellular matrix surrounding these endothelial cells, matrix metalloproteinases (MMPs) may be produced. The MMPs are a family of enzymes that degrade the extracellular matrix and include the collagenases, gelatinases A and B (72 kDa and 92 kDa gelatinases, respectively) and the stromelysins. The MMPs must be activated by proteolytic cleavage in vivo or by organomercurials or sodium dodecyl sulfate (SDS) in vitro. We wished to determine if VEGF could alter in vitro MMP production by human microvascular EC. Methods: Commercial microvascular EC isolated from human lungs (hMVEC-L; Clonetics: San Diego) were grown to confluency. Cells were plated on plastic cell culture ware with 10 without 10 ng/ml VEGF. Media covering the cells were removed after 24 hr and aralyzed by zymography to determine MMP production. Results: The control (untreated) hMVEC-L produced a gelatinase with an apparent Mr of 63 kDa. This enzyme is similar in molecular weight to activated gelatinase A and may represent the result of SDS activation in vitro. Treatment of an equal number of hMVEC-L with 10 ng/ml VEGF resulted in an increase in gelatinolytic activity by at least 100{\%}, indicating the presence of more gelatinase A. Conclusions: VEGF activation of microvascular endothelial cell proliferation is coordinated with an increase in gelatinase A production by these cells, indicating a possible mechanism for the coordination in proliferation and migration during PDR and other retinal pathologies that exhibit neovascularization.",
author = "Lamoreaux, {W. J.} and Fitzgerald, {M. E C} and Anton Reiner and Karen Hasty and Charles, {S. T.}",
year = "1996",
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T1 - Vascular Endothelial Growth Factor (VEGF) increases matrix metalloproteinase production by human microvascular endothelial cells in vitro

AU - Lamoreaux, W. J.

AU - Fitzgerald, M. E C

AU - Reiner, Anton

AU - Hasty, Karen

AU - Charles, S. T.

PY - 1996/2/15

Y1 - 1996/2/15

N2 - Purpose: Proliferation and migration of vascular endothelial cells (EC) are characteristic of retinal neovascularization during proliferative diabetic retinopathy (PDR). Proliferation of EC has been shown to be induced by vascular endothelial growth factor (VEGF). To facilitate migration through the extracellular matrix surrounding these endothelial cells, matrix metalloproteinases (MMPs) may be produced. The MMPs are a family of enzymes that degrade the extracellular matrix and include the collagenases, gelatinases A and B (72 kDa and 92 kDa gelatinases, respectively) and the stromelysins. The MMPs must be activated by proteolytic cleavage in vivo or by organomercurials or sodium dodecyl sulfate (SDS) in vitro. We wished to determine if VEGF could alter in vitro MMP production by human microvascular EC. Methods: Commercial microvascular EC isolated from human lungs (hMVEC-L; Clonetics: San Diego) were grown to confluency. Cells were plated on plastic cell culture ware with 10 without 10 ng/ml VEGF. Media covering the cells were removed after 24 hr and aralyzed by zymography to determine MMP production. Results: The control (untreated) hMVEC-L produced a gelatinase with an apparent Mr of 63 kDa. This enzyme is similar in molecular weight to activated gelatinase A and may represent the result of SDS activation in vitro. Treatment of an equal number of hMVEC-L with 10 ng/ml VEGF resulted in an increase in gelatinolytic activity by at least 100%, indicating the presence of more gelatinase A. Conclusions: VEGF activation of microvascular endothelial cell proliferation is coordinated with an increase in gelatinase A production by these cells, indicating a possible mechanism for the coordination in proliferation and migration during PDR and other retinal pathologies that exhibit neovascularization.

AB - Purpose: Proliferation and migration of vascular endothelial cells (EC) are characteristic of retinal neovascularization during proliferative diabetic retinopathy (PDR). Proliferation of EC has been shown to be induced by vascular endothelial growth factor (VEGF). To facilitate migration through the extracellular matrix surrounding these endothelial cells, matrix metalloproteinases (MMPs) may be produced. The MMPs are a family of enzymes that degrade the extracellular matrix and include the collagenases, gelatinases A and B (72 kDa and 92 kDa gelatinases, respectively) and the stromelysins. The MMPs must be activated by proteolytic cleavage in vivo or by organomercurials or sodium dodecyl sulfate (SDS) in vitro. We wished to determine if VEGF could alter in vitro MMP production by human microvascular EC. Methods: Commercial microvascular EC isolated from human lungs (hMVEC-L; Clonetics: San Diego) were grown to confluency. Cells were plated on plastic cell culture ware with 10 without 10 ng/ml VEGF. Media covering the cells were removed after 24 hr and aralyzed by zymography to determine MMP production. Results: The control (untreated) hMVEC-L produced a gelatinase with an apparent Mr of 63 kDa. This enzyme is similar in molecular weight to activated gelatinase A and may represent the result of SDS activation in vitro. Treatment of an equal number of hMVEC-L with 10 ng/ml VEGF resulted in an increase in gelatinolytic activity by at least 100%, indicating the presence of more gelatinase A. Conclusions: VEGF activation of microvascular endothelial cell proliferation is coordinated with an increase in gelatinase A production by these cells, indicating a possible mechanism for the coordination in proliferation and migration during PDR and other retinal pathologies that exhibit neovascularization.

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