Epidermal growth factor and laminin receptors contribute to migratory and invasive properties of gliomas.

Invasion & metastasis Pub Date : 1997-01-01
B B Tysnes, H K Haugland, R Bjerkvig
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Abstract

Gliomas are characterized by their extensive invasion into the brain parenchyma. Recently it has been shown that normal brain cells can produce laminin, fibronectin and collagen type IV when confronted by invading glioma cells. Laminin stimulates cell migration of several human glioma cell lines in vitro. This migration can be inhibited by adding blocking monoclonal antibodies (MAbs) against the most expressed integrin subunits, alpha3 and beta1. Previous studies have shown that glioma cell migration, invasion and growth are stimulated by epidermal growth factor (EGF). However, MAb directed against the EGF receptor (EGFR) did only partly inhibit the invasive process in vitro. Since laminin has regional peptide homology with EGF (EGF-like repeats), the present work was aimed at studying how two human glioma cell lines exposed to antibodies to the EGFR, reacted to laminin stimulated migration. Furthermore, we wanted to study which role the EGFR and the laminin receptor integrin subunits alpha3 and beta1 play during glioma cell invasion. EGFR expression of two glioma cell lines, AN1/lacZ and U-251/lacZ was studied by flow cytometry and immunofluorescence microscopy. A cell migration assay was used to study effects of MAbs against EGFR on migration from laminin-stimulated tumor spheroids. Tumor cell invasion was evaluated by using an in vitro co-culture model, where normal fetal brain cell aggregates were confronted with multicellular tumor spheroids. The results show that both cell lines expressed EGFR, AN1/lacZ 4-fold more than U-251/lacZ. MAb against EGFR inhibited the laminin-stimulated migration only from AN1/lacZ spheroids. MAbs against alpha3 and beta1 integrin subunits inhibited glioma cell invasion in vitro. The present work indicates possible connections between laminin-stimulated cell migration and the EGFR expression on glioma cells. These elements contribute to the characteristic features of glioma cells and may be an important part of the complex relationships between growth factors, integrins and extracellular matrix during glioma cell invasion.

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表皮生长因子和层粘连蛋白受体参与胶质瘤的迁移和侵袭特性。
胶质瘤的特点是广泛侵袭脑实质。最近有研究表明,正常脑细胞在面对入侵的胶质瘤细胞时,可以产生层粘连蛋白、纤维连接蛋白和IV型胶原蛋白。层粘连蛋白刺激几种人类胶质瘤细胞系的细胞迁移。这种迁移可以通过添加针对最表达的整合素亚单位alpha3和beta1的阻断单克隆抗体(mab)来抑制。已有研究表明,表皮生长因子(epidermal growth factor, EGF)可促进胶质瘤细胞的迁移、侵袭和生长。然而,针对EGF受体(EGFR)的单抗在体外仅部分抑制了侵袭过程。由于层粘连蛋白与EGF具有区域肽同源性(EGF样重复序列),目前的工作旨在研究暴露于EGFR抗体的两种人类胶质瘤细胞系如何对层粘连蛋白刺激的迁移做出反应。此外,我们希望研究EGFR和层粘连蛋白受体整合素亚基α 3和β 1在胶质瘤细胞侵袭过程中发挥的作用。用流式细胞术和免疫荧光显微镜研究了两种胶质瘤细胞株AN1/lacZ和U-251/lacZ的EGFR表达。细胞迁移试验用于研究抗EGFR单克隆抗体对层粘连蛋白刺激的肿瘤球体迁移的影响。肿瘤细胞侵袭通过体外共培养模型进行评估,其中正常胎儿脑细胞聚集体与多细胞肿瘤球体相对抗。结果表明,两种细胞系表达EGFR, AN1/lacZ均比U-251/lacZ高4倍。抗EGFR单克隆抗体仅抑制层粘连蛋白刺激的AN1/lacZ球体的迁移。针对α 3和β 1整合素亚基的单克隆抗体体外抑制胶质瘤细胞侵袭。本研究提示层粘连蛋白刺激的细胞迁移与胶质瘤细胞上EGFR的表达之间可能存在联系。这些元素有助于胶质瘤细胞的特征,可能是胶质瘤细胞侵袭过程中生长因子、整合素和细胞外基质之间复杂关系的重要组成部分。
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