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Interaction with fibronectin regulates cytokine gene expression in human melanoma cells

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.