Research ArticleLaminin isoforms and their integrin receptors in glioma cell migration and invasiveness: Evidence for a role of α5-laminin(s) and α3β1 integrin
Introduction
Gliomas are the most common primary brain tumors and are often highly malignant. Glioblastoma multiforme, the most aggressive glioma, has a poor prognosis and most patients die of their disease in less than a year, independently of treatment [1]. Malignant glioma diffusely infiltrates the normal brain tissue as single cells, and this infiltration makes complete surgical resection practically impossible. The invasive cells often migrate along myelinated fiber tracts and in the perivascular space along blood vessels [2], [3], [4], [5]. In the latter case, the glioma cells interact with the abluminal side of the vessels, adopting a pericytic-like location, without intravasating. This pattern of migration may be due to specific interactions between extracellular matrix (ECM) components of the vascular basement membrane (BM) and tumor cell surface receptors. Alternatively, glioma tumors may create a permissive migratory substrate by synthesizing and depositing autologous ECM components.
Laminins are a growing family of large heterotrimeric αβγ proteins which promote cell adhesion and migration via integrin receptors [6], [7], [8]. Together with type IV collagen, perlecan and nidogen, these ECM proteins are major components of all BMs, including the vascular BM [9]. Five α, three β and three γ chains constitute by combination over 15 different laminin isoforms, and the α chains, which display a cell- and tissue-specific expression, are differentially recognized by nearly 10 out of 24 different integrins [6], [10]. In a new simplified nomenclature, laminin isoforms are designated according to their chain composition (e.g., laminin-8 or α4β1γ1 is now called Lm-411) [8]. Though the first laminin, Lm-111 or laminin-1, was reported over 25 years ago, a large number of additional laminin isoforms with wider tissue distribution, different integrins receptors and different biological activities have been identified since then.
Several studies concerning gliomas and laminins have been reported in the literature. By immunohistochemistry, laminin has been localized mainly to the tumor vasculature, but also within the tumor as punctate deposits and at the brain/tumor confrontation zone, particularly in glioma models [11], [12], [13], [14]. Glioma cells are known to secrete several laminins, but their isoforms are poorly characterized [15], [16]. Moreover, among several BM components and other ECM proteins, laminins appeared to be most permissive for adhesion and migration of glioma cells in vitro, and these interactions were mediated by integrins, including α3β1 and α6β1 [16], [17], [18], [19], [20], [21], [22], [23]. Compared to normal brain and astrocytes, astrocytomas and glioblastomas showed increased expression of the former integrin both in vivo and in vitro [24]. These studies have provided most valuable information. However, the anti-laminin antibodies used were often cross-reactive with most laminin isoforms, or their chain specificity was unknown. Moreover, the laminin preparations were often poorly characterized molecularly, differing from one another and consisting of highly fragmented proteins, and/or a mixture of laminin isoforms [25], [26]. In addition, only a limited number of laminin isoforms, predominantly Lm-111, or their integrin receptors were tested, and no systematic analysis of α5-laminin(s) was reported.
More recently, the chain specificity of many laminin antibodies has been established, and several recombinant laminin isoforms have been produced [25], [26], [27], [28], [29]. In the present study, a panel of antibodies to all laminin chains, except γ3, was used to identify the laminins of glioma tumor tissue, particularly of the vasculature. Moreover, the glioma cell migration-promoting activity of laminins covering all known α chains was established by using fully characterized natural and/or recombinant proteins, and their major integrin receptor was identified with a panel of antibodies to all known integrins. Finally, the effect of blocking antibodies to this integrin in the invasiveness of human glioma cells transplanted into the brain of nude mice was determined, and the laminin isoforms secreted by glioma cells were identified.
Section snippets
Cells, purified proteins and mAbs
Human malignant glioma cell lines (A172, KG1C, T98G, U251) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum, 2 mmol/L glutamine, and antibiotics. All cell lines were obtained from the Health Science Research Resources Bank, Tokyo, Japan. α1-Laminin (Lm-111, laminin-1) was isolated from mouse Engelbreth–Holm–Swarm (EHS) tumor and obtained from Chemicon (Chemicon International, Temecula, CA). α2-Laminins were used either as merosin isolated from human placenta (Lm-211
Glioma tissue vasculature expresses α2-, α3-, α4- and α5-laminins
To identify the laminin isoforms in the vasculature of gliomas, immunohistochemistry of glioblastoma tissues was first performed with mAbs to laminin chains (Fig. 1). The tumor vessels were strongly stained with mAbs to Lmα2, Lmα4, Lmα5, Lmβ1 and Lmγ1, whereas staining for Lmα3 and Lmβ2 was less intense. Faint or no staining was observed with mAbs to Lmα1, Lmβ3 and Lmγ2, though the mAb to Lmα1 clearly reacted with the vasculature of normal brain tissue (Fig. 1 and data not shown). Under the
Discussion
In the present study, the migration-promoting activity of laminins covering all known α chains was systematically compared on glioma cells for the first time, and a laminin isoform-specific effect was observed. α3- and α5-laminins, which were found in the tumor vasculature, strongly promoted migration, whereas α2- and α4-laminins exerted a minimal effect, if any. α1-Laminin displayed an intermediate activity. Among all integrins, α3β1 was identified as the most abundant in glioma cells, and as
Note added in proof
During revision of the present paper, Chia et al. (Am J Pathol 170:2135 2007) reported evidence for a role of tumor-derived laminin-511 in the metastatic progression of breast cancer. Moreover, additional studies from our laboratory have recently demonstrated expression, recognition and use of α5-laminin(s) by human malignant melanoma cells (Oikawa and Patarroyo, unpublished data), similarly to glioma cells. These findings indicate participation of α5-laminin(s) and α3β1 integrin in several
Acknowledgments
The authors thank Drs. Ismo Virtanen, Eva Engvall, Kiyotoshi Sekiguchi, Randall Kramer and Samuel Wright for providing mAbs, Drs. Sergei Smirnov and Peter Yurchenco for providing rLm-211, and Drs. Monica Nistér and Bengt Westermark for their comments on the manuscript. This work was supported by the Swedish Cancer Society and Karolinska Institutet.
References (55)
- et al.
Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion
Int. J. Biochem. Cell Biol.
(2004) - et al.
Laminin isoforms in tumor invasion, angiogenesis and metastasis
Semin. Cancer Biol.
(2002) - et al.
A simplified laminin nomenclature
Matrix Biol.
(2005) Integrins: bidirectional, allosteric signaling machines
Cell
(2002)- et al.
Structural analysis of a human glial variant laminin
Exp. Cell Res.
(1996) - et al.
Purification and characterization of human laminin-8. Laminin-8 stimulates cell adhesion and migration through alpha3beta1 and alpha6beta1 integrins
J. Biol. Chem.
(2001) - et al.
The neurite-promoting domain of human laminin promotes attachment and induces characteristic morphology in non-neuronal cells
Exp. Cell Res.
(1988) - et al.
Chain specificity assignment of monoclonal antibodies to human laminins by using recombinant laminin beta1 and gamma1 chains
Matrix Biol.
(2000) - et al.
Characterization of commercial laminin preparations from human placenta in comparison to recombinant laminins 2 (alpha2beta1gamma1), 8 (alpha4beta1gamma1), 10 (alpha5beta1gamma1),
Matrix Biol.
(2006) - et al.
Recombinant human laminin-10 (alpha5beta1gamma1). Production, purification, and migration-promoting activity on vascular endothelial cells
J. Biol. Chem.
(2002)
Chondroitin sulphate modification in the alpha4 chain of human recombinant laminin-8 (alpha4beta1gamma1)
Matrix Biol.
Contributions of the LG modules and furin processing to laminin-2 functions
J. Biol. Chem.
An endothelial laminin isoform, laminin 8 (alpha4beta1gamma1), is secreted by blood neutrophils, promotes neutrophil migration and extravasation, and protects neutrophils from apoptosis
Blood
Laminin chain expression suggests that laminin-10 is a major isoform in the mouse hippocampus and is degraded by the tissue plasminogen activator/plasmin protease cascade during excitotoxic injury
Neuroscience
Downregulation of major histocompatibility complex antigens in invading glioma cells: stealth invasion of the brain
Lab. Invest.
Rac regulates integrin-mediated endothelial cell adhesion and migration on laminin-8
Exp. Cell Res.
The role of integrin receptors in aspects of glioma invasion in vitro
Int. J. Dev. Neurosci.
Ligand-binding specificities of laminin-binding integrins: a comprehensive survey of laminin-integrin interactions using recombinant alpha3beta1, alpha6beta1, alpha7beta1 and alpha6beta4 integrins
Matrix Biol.
Structural requirement of carboxyl-terminal globular domains of laminin alpha 3 chain for promotion of rapid cell adhesion and migration by laminin-5
J. Biol. Chem.
The LG3 module of laminin-5 harbors a binding site for integrin alpha3beta1 that promotes cell adhesion, spreading, and migration
J. Biol. Chem.
Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10
J. Biol. Chem.
Domain IVa of laminin alpha5 chain is cell-adhesive and binds beta1 and alphaVbeta3 integrins through Arg-Gly-Asp
FEBS Lett.
Transfection of MCF-7 carcinoma cells with human integrin alpha7 cDNA promotes adhesion to laminin
Arch. Biochem. Biophys.
Brain tumors
Pericytic-like angiotropism of glioma and melanoma cells
Am. J. Dermatopathol.
Mechanisms of tumor cell invasion and angiogenesis in the central nervous system
Front. Biosci.
Transplanted glioma cells migrate and proliferate on host brain vasculature: a dynamic analysis
Glia
Cited by (86)
Exploring the Vital Link Between Glioma, Neuron, and Neural Activity in the Context of Invasion
2023, American Journal of PathologyQuantification of glioblastoma progression in zebrafish xenografts: Adhesion to laminin alpha 5 promotes glioblastoma microtumor formation and inhibits cell invasion
2018, Biochemical and Biophysical Research CommunicationsThe role of CD146 in renal disease: from experimental nephropathy to clinics
2024, Journal of Molecular MedicineEvaluating glioblastoma tumour sphere growth and migration in interaction with astrocytes using 3D collagen-hyaluronic acid hydrogels
2023, Journal of Materials Chemistry B