Immunohistochemical analysis of reactive astrocytes around glioblastoma: an immunohistochemical study of postmortem glioblastoma cases

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Abstract

To investigate the mechanisms of proteolysis within the glioma, and tissue reactions against glioblastoma, immunohistochemical detection both outside and inside of the tumor was performed using seven brains with glioblastoma that were obtained from autopsies. Immunohistochemistry was performed to detect vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-1,-2,-9, membrane-type matrix metalloproteinase (MT-MMP), interleukin (IL)1-β, and IL-6. The data were translated into color graphics and the localization of these proteins was analyzed. In glial cells around the tumor, GFAP, VEGF, MMP-2, and MT-MMP were strongly expressed. Moreover, IL1-β was also expressed strongly in the glial cells at the periphery of the tumor. IL-6 was recognized outside of the tumor, but was expressed only in the swollen astrocytes and normal pyramidal cells. These data suggest that in the periphery of the tumor, tissue reconstruction processes take place with concomitant degradation of the matrix by MMP-2 and MT-MMP, as well as vascular remodeling promoted by VEGF. The fact that IL1-β, but not IL-6, was expressed strongly in the glial cells around the tumor, may indicate that these proteins expressed outside of the tumor are not utilized for tumor growth, but may be used to guard the tumor against invasions, such as immune response.

Introduction

Tumor cell invasion requires proteolytic degradation of extracellular matrices and neovascularization in the peripheral area adjacent to the tumor [1], [2]. Plasminogen activators and proteins of the matrix metalloproteinase (MMP) family are the major proteases that degrade peritumoral extracellular matrices. Vascular endothelial growth factor (VEGF) is a protein important for vascular proliferation within the tumor.

MMPS have been implicated in proteolytic degradation processes. Secretion of MMPs by tumor cells has been demonstrated in malignant brain tumors, and reportedly is associated with malignant progression of these neoplasms [3]. Reportedly, malignant gliomas express MMP-1, MMP-2, MMP-9 and TTMP-1 [4], [5]. In addition, vascular endothelial growth factor (VEGF) is normally active in embryogenesis and wound healing, and also is important in the growth of various tumors, including glioblastoma [6], [7], [8], [9], [10], [11].

On the other hand, proteolytic degradation and neovascularization outside of the tumor must occur not only in association with the tumor cell invasion, but also to facilitate tissue reconstruction processes that guard against the tumor cell invasion.

In this study, which entails immunohistochemistry on seven autopsy brain specimens with glioblastoma, we investigated the roles of MMPs, and VEGF in glioma cell invasion to the adjacent structures, and anti-tumor responses of adjacent brain tissues using graphic analysis.

Section snippets

Tissue preparation

Seven autopsy brain specimens from patients who died of glioblastoma, including four males and three females, were used for this study. The average age at diagnosis was 67.3 years. Patient's ages were in a range from 26 to 86 years old and the survival period after diagnosis was 480±502.7 days (mean±S.D.). Six patients were treated with surgical interventions followed by radiotherapy (average, 55 Gy) and intravenous infusion of 1-(4-amino-2-methyl-5-pyrimidinyl)

Results

Fig. 1 shows the result of MRI T1-Gd, where the patient was examined 1 month prior to death. Ki-67 and VEGF immunopositivity are also shown color graphically in a slice of whole brain. As demonstrated here, Ki-67 positive cells are mainly localized in the tumor, however, VEGF immunopositivity was stronger outside the tumor than in the tumor itself. This phenomenon strongly suggests that there is a natural tissue reaction against the tumor.

Fig. 2 demonstrates the immunopositivity of each protein

Discussion

The mechanisms by which glioma cells migrate and invade adjacent normal brain tissue are still under investigation, but cell-matrix proteolysis and neovascularization are necessary elements. In these processes, MMPs have proven to be key enzymes for glioma invasion. Among the 11 known MMPs, MMP-2 and -9 have been found to be associated with tumor cell invasion. The activated forms of MMP such as MT-MMP are necessary for proteolytic activity, and MMP function is influenced by antiproteolytic

References (25)

  • G. Nagashima et al.

    Graphic analysis of microscopic tumor cell infiltration, proliferative potential, and vascular endothelial growth factor expression in an autopsy brain with glioblastoma

    Surg. Neurol.

    (1999)
  • M.E. Smith et al.

    Differential effect of interleukin-1 beta on Ia expression in astrocytes and microglia

    J. Neuroimmunol.

    (1993)
  • L.M. Matrisian

    The matrix-degradation metalloproteinases

    Bioassays

    (1992)
  • P. Mignatti et al.

    Biology and biochemistry of proteinases in tumor invasion

    Physiol. Rev.

    (1993)
  • J.S. Rao et al.

    Elevated levels of Mr 92000 type IV collagenase in human brain tumors

    Cancer Res.

    (1993)
  • G. Apodaca et al.

    Expression of metalloproteinases and metalloproteinase inhibitors by fatal astrocytes and glioma cells

    Cancer Res.

    (1990)
  • T. Nakagawa et al.

    Production of matrix metalloproteinases and tissue inhibitor of metalloproteinases-1 by human brain tumors

    J. Neurosurg.

    (1994)
  • R.A. Berkman et al.

    Expression of the vascular permeability factor/vascular endothelial growth factor gene in central nervous system neoplasms

    J. Clin. Invest.

    (1993)
  • G. Breier et al.

    Expression of vascular endothelial growth-factor during embryonic angiogenesis and endothelial-cell differentiation

    Development

    (1992)
  • L.F. Brown et al.

    Expression of vascular permeability factor (vascular endothelial growth factor) by epidermal keratinocytes during wound healing

    J. Exp. Med.

    (1992)
  • E. Hashimoto et al.

    Rapid induction of vascular endothelial growth factor expression by transient ischemia in rat heart

    Am. J. Physiol.

    (1994)
  • H.C. Hollingsworth et al.

    Tumor angiogenesisin in advanced stage ovarian carcinoma

    Am. J. Pathol.

    (1995)
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