Abstract
Our study was aimed at investigating whether interaction of human melanoma cells with the extracellular matrix (ECM) protein fibronectin (FN) could regulate lymphokine gene expression. Serum-deprived cells (quiescent condition) of a metastatic melanoma cloned line were cultured either on uncoated or on FN- or BSA-coated surfaces. By means of reverse transcriptase-polymerase chain reaction (RT-PCR), we analyzed mRNA expression of 4 cytokines—interleukin (IL)-Iα, IL-Iβ, IL-6 and IL-8—and 9 growth factors—endothelial cell growth factor (ECGF), basic fibroblast growth factor (bFGF), fibroblast growth factor (FGF)-5, HST, keratinocyte growth factor (KGF), transforming growth factor (TGF)-α, TGF-β1, TGF-β2 and TGF-β3. When cultured on FN, melanoma cells expressed IL-Iα and IL-6 transcripts in addition to IL-Iβ, IL-8, ECGF, TGF-β1, TGF-β2 and TGF-β3, already present in quiescent cells. Amplification parameters to achieve semi-quantitative RT-PCR were then determined for each detectable factor, thus allowing us to measure a selective enhancement of mRNA levels for IL-1α, IL-6, IL-8 and TGF-β2 upon interaction with FN by quiescent melanoma cells. This augmented expression was inhibited by an anti-integrin β1 chain monoclonal antibody (MAb). Moreover, the amounts of IL-6, IL-8 and IL-β produced in the supernatants, as assessed by ELISA, correlated with the corresponding mRNA expression. Extension of this analysis to the other 5 human primary and metastatic melanoma lines confirmed the ability of FN to selectively up-regulate only IL-6 and IL-8 secretion. Our data indicate that FN is able to modulate expression and secretion of a defined subset of lymphokines in human melanoma. © 1996 Wiley-Liss, Inc.
Published Version
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