Fibronectin-mediated cell adhesion is required for induction of 92-kDa type IV collagenase/gelatinase (MMP-9) gene expression during macrophage differentiation. The signaling role of protein kinase C-beta.
Induction of the 92-kDa gelatinase (MMP-9) gene expression is associated with macrophage differentiation. In this study, we explored the regulatory mechanisms underlying this differentiation-associated MMP-9 gene expression in human HL-60 myeloid leukemia cells and human peripheral blood monocytes. Phorbol 12-myristate 13-acetate (PMA) markedly induced MMP-9 gene expression in HL-60 cells; the induction closely paralleled the timing and extent of PMA-induced cell adhesion and spreading, a hallmark of macrophage differentiation. Similarly, treatment with PMA or macrophage-colony stimulating factor stimulated adherence and spreading of blood monocytes with a concurrent 7- or 5-fold increase in MMP-9 production, respectively. In protein kinase C (PKC)-beta-deficient HL-60 variant cells (HL-525), PMA failed to induce cell adhesion and MMP-9 gene expression. Transfecting HL-525 cells with a PKC-beta expression plasmid restored PKC-beta levels and PMA inducibility of cell adhesion and spreading as well as MMP-9 gene expression. Induction of cell adhesion and MMP-9 gene expression in HL-60 cells and blood monocytes was strongly inhibited by neutralizing monoclonal antibodies to fibronectin (FN) and its receptor alpha5 beta1 integrin. HL-525 cells, which constitutively display high levels of surface alpha5 beta1 integrin, adhered and spread on immobilized FN with concomitant induction of MMP-9 gene expression. Cytochalasins B and D were each a potent inhibitor of MMP-9 production. Our results suggest that alpha5 beta1 integrin-mediated interaction of immature hematopoietic cells with FN plays a critical role in modulating matrix-degrading activities during macrophage differentiation.
Xie
B
B
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, Argonne, Illinois 60439-4833, USA.
Laouar
A
A
Huberman
E
E
D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects; D017364:Collagenases / Q000096:biosynthesis*; D003571:Cytochalasin B / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D005353:Fibronectins / Q000502:physiology; D018922:HL-60 Cells; D006801:Humans; D007527:Isoenzymes / Q000502:physiology*; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008264:Macrophages / Q000201:enzymology*; D020780:Matrix Metalloproteinase 9; D009000:Monocytes / Q000166:cytology / Q000201:enzymology*; D011493:Protein Kinase C / Q000502:physiology*; D064546:Protein Kinase C beta; D017469:Receptors, Fibronectin / Q000502:physiology; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology
D005353:Fibronectins; D007527:Isoenzymes; D017469:Receptors, Fibronectin; D003571:Cytochalasin B; D014212:Tretinoin; D016173:Macrophage Colony-Stimulating Factor; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D017364:Collagenases; D020780:Matrix Metalloproteinase 9; D013755:Tetradecanoylphorbol Acetate