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Characterisation of ligand binding to the parathyroid hormone/parathyroid hormone‐related peptide receptor in MCF7 breast cancer cells and SaOS‐2 osteosarcoma cells
Author(s) -
Alokail Majed S.,
Peddie Margaret J.
Publication year - 2005
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.1280
Subject(s) - parathyroid hormone , medicine , endocrinology , parathyroid hormone related protein , receptor , calcitriol , chemistry , ligand (biochemistry) , osteosarcoma , parathyroid hormone receptor , cancer cell , calcitriol receptor , hormone receptor , cancer research , biology , calcium , breast cancer , cancer
Parathyroid hormone‐related peptide (PTHrP) and parathyroid hormone (PTH)/PTHrP‐receptor, PTH/PTHrP‐R, are frequently expressed in mammary carcinomas as well as in bone cells. In this study we compared the ligand binding characteristics of the PTH/PTHrP–R in SaOS‐2 human osteosarcoma cells with those in MCF7 breast cancer cells. We used both Scatchard analysis of saturation kinetics for iodinated ligand and the level of expressed receptor protein by visualising the single radio‐labelled receptor‐ligand complex from isolated membrane preparations from the two cell lines. In MCF7 cells, ligand binding, (receptor number) was increased by prior exposure of the cultured cells to epidermal growth factor (EGF), estradiol (E2), or dexamethasone (DEX), and decreased following calcitriol (1,25 DHCC). In contrast in the SaOS‐2 cells, PTH/PTHrP‐R number was increased by exposure to E2 and 1,25DHCC and decreased by DEX while EGF had no effect. These data were confirmed when the PTH/PTHrP‐R was cross linked with 125 I‐PTHrP‐1‐34 Tyr , and extended by visualising the intensity of the isolated radiolabelled receptor complex by autoradiography following SDS‐PAGE at several time points during the treatment. Copyright © 2005 John Wiley & Sons, Ltd.