ErbB receptors mediate both migratory and proliferative activities in human melanocytes and melanoma cells

Abstract

Epidermal growth factor receptor (EGFR) activation by transforming growth factor alpha (TGFalpha) has been implicated in autocrine growth in melanoma, but does not alter melanocyte proliferation. This raises the possibility that different signalling pathways are activated via EGFR or ErbB receptors. Here, we demonstrate that ErbB2, ErbB3 and ErbB4 are expressed in cultured human melanocytes. Western analyses with receptor-specific antisera revealed protein bands with Mr values of 185 and 160 kDa, corresponding to ErbB2 and ErbB3, respectively. Blots probed with ErbB4 antibodies showed bands with Mr values of 180, 120 and 80 kDa, corresponding to the receptor and its reported variants. Two malignant melanoma cell lines expressed ErbB2 and ErbB3, but not the full-length ErbB4 receptor. As TGFalpha binds to EGFR and the heregulins (HRG) bind to ErbB3 and ErbB4, these growth factors were examined for effects on receptor activation and on cell growth and motility in a scratch wound closure assay. In normal melanocytes, HRGbeta1 activated the phosphorylation of tyrosine residues of proteins that immunoprecipitated with EGFR and ErbB4 antisera, and significantly enhanced cell migration but not proliferation. Neither TGFalpha nor HRGalpha1 promoted migration or growth in normal melanocytes. By contrast, TGFalpha stimulated migratory activity in the MM96L cell line, but not in the MELJG line, whereas HRGbeta1 significantly enhanced cell growth, but not migration, in both malignant cell lines. The apparent transition of HRGbeta1 from a migratory to a proliferative function after malignant transformation, and the change in TGFalpha from a non-migratory to a migratory activity in one melanoma line, suggests multiple switches in ErbB signalling pathways via EGFR/ErbB heterodimer formation.