Experimental hypoxia does not influence Eph receptor and ephrin ligand expression in human melanoma cell lines

Experimental evidence links the receptor tyrosine kinases EphA2 and EphB4 to melanoma progression and metastasis. Another factor contributing to melanoma invasion-metastasis cascade is hypoxia. However, data on the influence of tumor hypoxia on regulation of Eph receptors and their ephrin ligands are scarce. This study aimed at clarifying whether hypoxia influences expression and synthesis of EphA2 and EphB4 and, furthermore, ephrinA1 and ephrinB2, respectively, in four human melanoma cell lines (A375, A2058, MeWo, and MelJuso). In order to investigate Eph/ephrin expression under hypoxic conditions we used cell monolayer cultures as extrinsic hypoxia models and A2058 spheroids as intrinsic hypoxia model. Hypoxic conditions were approved by measurement of VEGF expression and cellular uptake of [18F]fluoromisonidazol ([18F]FMISO). In all models, both VEGF expression and ([18F]FMISO) uptake increased under hypoxic conditions. In normoxia, EphA2, EphB4, ephrinA1, and ephrinB2 expression was detectable in all cell lines showing a substantially varying extent. Protein synthesis of EphA2 was detected in all cell lines and, moreover, of EphB4 in A375 and A2058 cells. However, no effect of experimental hypoxia on Eph/ephrin expression and protein synthesis could be observed. The findings contribute to debating the hypothesis that hypoxia is an important regulator of Eph/ephrin expression in tumors.