Integrins induce expression of monocyte chemoattractant protein-1 via focal adhesion kinase in mesangial cells

Background. Integrins are major adhesion receptors that not only regulate cytoskeletal organization, but also trigger a variety of intracellular signal transduction pathways. We examined the effects of increased extracellular matrix (ECM) accumulation on monocyte chemoattractant protein-1 (MCP-1) expression, which is known to play an important role in the progression of various glomerular diseases. Methods. MCP-1 mRNA and protein expression in cultured rat mesangial cells (MC) attached to ECM proteins were examined by reverse transcription (RT)-polymerase chain reaction (PCR) and Western blotting, respectively. Phosphorylation of focal adhesion kinase (FAK) was measured by Western blotting. Effects of wild-type and dominant-negative FAK on MCP-1 expression were examined by a transient transfection assay. Results. Cell adhesion to fibronectin-induced phosphorylation of FAK and MCP-1 mRNA expression in time- and dose-dependent manners followed by increased MCP-1 protein expression. All integrin-interacting substrates (laminin and types I, III, and IV collagens) also increased levels of FAK phosphorylation and MCP-1 expression, whereas nonspecific adhesive substrates (polylysine and concanavalin A) had no significant effects. Overexpression of wild-type FAK increased phosphorylation of FAK and expression of MCP-1 mRNA and protein, whereas transfection of dominant-negative FAK abolished adhesion-induced MCP-1 expression. Adhesion-induced expression of MCP-1 mRNA was inhibited by genistein and tosyl phenylalanyl chloromethylketone (TPCK), suggesting that tyrosine kinases [e.g., FAK, and nuclear factor kappa B (NF-kappaB)] are necessary in this signaling. Conclusion. Our results indicate that integrin-mediated cell adhesion to the ECM can induce MCP-1 expression through activation of FAK, and suggest a role for altered ECM deposition in the progression of glomerular diseases by affecting gene expression.