C. Bagutti et al., ALPHA-MSH RECEPTOR AUTORADIOGRAPHY ON MOUSE AND HUMAN-MELANOMA TISSUE-SECTIONS AND BIOPSIES, Journal of receptor and signal transduction research, 15(1-4), 1995, pp. 427-442
MSH receptors and their binding characteristics of [I-125]-labelled de
rivatives of alpha-MSH have been studied extensively on various mouse
and human melanoma cell lines in culture. The aim of this study was to
determine the binding characteristics of alpha-MSH radioligands to MS
H receptors occurring in experimental mouse and human melanoma tumours
as well as in human melanoma biopsies. For this reason, solid tumours
were grown on experimental animals by inoculation of murine B16-F1 an
d human D10 and HBL melanoma cells. After excision and cryosectioning
of the tumours, frozen tissue sections were incubated with [(I-125)Tyr
(2)]-alpha-MSH or [(I-I25)Tyr(2),Nle(4),D-Phe(7)]-alpha-MSH and specif
ic alpha-MSH binding sites were visualized by subsequent autoradiograp
hy. The presence of increasing concentra-tions of unlabelled alpha-MSH
during incubation with tracer led to a dose-dependent displacement of
the radioligand. Quantitative analysis of the autoradiograms produced
dissociation constants which were comparable with those obtained with
cell binding assays: K-D = 1.87 and 1.31 nmol/l for B16 tumours and c
ells, respectively; 0.32 and 0.33 nmol/l for D10, and 2.24 and 1.36 nm
ol/l for HBL tumours and cells, respectively. This indicates similar b
inding properties of alpha-MSH radioligands to both cultured melanoma
cells and tissue sections of melanoma tumours from experimental animal
s. Similar binding characteristics were also observed with human melan
oma tissue sections originating from biopsies of melanoma patients.