Effects of collagen IV on neuroblastoma cell matrix-related functions.
Integrin-mediated interactions with collagen IV and its domains were examined in a human neuroblastoma cell line (SK-N-SH). By adhesion assays we demonstrated that neuroblastoma cells bound to solid-phase intact collagen IV and synthetic cell-binding peptide HEP-III, derived from the collagenous part of the molecule, but not to the main noncollagenous NC1 domain or to the synthetic cell-binding peptide HEP-I, derived from this domain. Monoclonal antibodies against beta1, alpha3, and alpha(v)beta3 integrins resulted in inhibition of cell binding to collagenous substrates by 95, 30, and 35%, respectively. By flow cytometry and immunoblotting it was shown that culture of SK-N-SH cells on collagen IV resulted in alteration in the expression of major neuroblastoma cell integrins. Binding to collagen IV induced the expression and activation of matrix metalloproteinases A and B (MMP-2, MMP-9), with a concomitant increase at the protein level of tissue-specific inhibitors of metalloproteinases (TIMP-1, TIMP-2). Finally, the expression of MMP-2 was significantly up-regulated by anti-alpha3beta1 antibodies, whereas ligation of anti-alpha(v)beta3 antibodies resulted in a modest down-regulation of MMP-2. Our results indicate that the presence of collagen IV modulates the expression of integrins, which are used for binding to this glycoprotein, and MMP-2 secreted by SK-N-SH cells.
Tzinia
Athina K
AK
Institute of Biology, National Center for Scientific Research "Demokritos,", 15310 Agia Paraskevi, Athens, Greece. atzin@mail.demokritos.gr
Kitsiou
Paraskevi V
PV
Talamagas
Argiris A
AA
Georgopoulos
Angelique
A
Tsilibary
Effie C
EC
D001386:Azides / Q000494:pharmacology; D001665:Binding Sites / Q000187:drug effects / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D024141:Collagen Type IV / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005109:Extracellular Matrix / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009447:Neuroblastoma; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000502:physiology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism; D014407:Tumor Cells, Cultured
D001386:Azides; D024141:Collagen Type IV; D003517:Cyclopentanes; D016023:Integrins; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; C058188:4,4-dimethyl-1,7-dioxa-2,6-diaza-7a-thia-3H,5H-benzo(cd)pentalene; C058190:1,7-(dioxa)-2,6-diaza-4,7a-dithia-3H,5H-benzo(cd)pentalene-4-oxide; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9