Transforming Growth Factor-β Stimulates the Expression of Fibronectin by Human Keratinocytes

abstractTransforming growth factor beta (TGF-β)is a 25-kD protein which has regulatory activity over a variety of cell types. It is distinct form epidermal growth factor (EGF) and EGF analogs, and exerts its action via a distinct receptor. Its effect on proliferation or differentiation can be positive or negative depending on the cell type and the presence of other growth factors. it also modulates the expression of cellular products. TGF-β causes fibroblasts to increase their production of the extracellular matrix components, fibronectin and collagen. human keratinocytes (HK) are known to have TGF-βreceptors. We wished to study the effect of TGF-β on the production of extracellular matrix proteins by human keratinocytes in culture.Human keratinocytes were grown in serum-free defined medium (MCDB-153) to about 70% confluence. Following a 16-h incubation in medium lacking EGF and TGF-β, cells were incubated for 12h in medium containing varying concentrations of EGF and TGF-β. Cells were then labeled with 35S-methionine for 10 h in the same conditions. Labeled proteins from the medium were analyzed by SDS-PAGE and autoradiography.TGF-β at 10 ng/ml induced a sixfold increase in the secretion of fibronectin, as well as an unidentified 50-kD protein. Thrombospondin production was also increased, but not over a generalized twofold increase in the production of all other proteins. EGF, at 10 ng/ml, caused a smaller additive effect, TGF-β may be an important stimulator of extracellular matrix production by human keratinocytes