Mannose-6-phosphate/insulin-like growth factor-II receptor in human melanoma cells: effect of ligands and antibodies on the receptor expression.

ABSTRACT

BACKGROUND: The M6P/IGF-II receptor belongs to the IGF system which plays a crucial role in tumorigenicity. While the role of the IGF-I receptor in signal transduction is well documented, previous experiments failed to uncover a clear signalling function for the M6P/IGF-II receptor. However, more recent studies have shown the capability of M6P/IGF-II receptor to initiate transmembrane signalling. MATERIALS AND METHODS: Human melanoma cells were used to detect the cell surface expression of the M6P/IGF-II receptor and its modulation by different effectors and monoclonal anti-receptor antibodies. RESULTS: M6P (5 mM) caused an increase of the luminescent receptor signal of about 50% . Pre-incubation of cells with Act-D (5 microg/mL) or CHI (10 microg/mL) following M6P stimulation in the presence of the inhibitors caused a reduction of receptor cell surface expression of 27% or 31%, respectively. The monoclonal antibody (mAb) 2G11 was able to mimic the M6P effect on the receptor up-regulation but the mAb MEM-238 did not. The synergistic effect detected with the combination of M6P and the mAb 2G11 and the failure of 2G11 to compete with the M6P action suggests that both effectors have different binding sites on the receptor. Unlike 2G11 the mAb MEM-238 prevented the M6P effect on receptor up-regulation confirming partially overlapping binding epitopes of both effectors. Brefeldin A was shown to have an inhibiting effect on the vesicular transport of the receptor protein to the plasma membrane and forskolin had an activating effect on the receptor exocytosis with the following enhanced integration into the plasma membrane. CONCLUSION: Up-regulation of the tumour suppressor M6P/IGF-II receptor might represent an approach for anticancer therapy. In addition, results support recent data on the receptor's capability of signal transduction.