Affinity isolation of cultured tumor cells by means of drugs and hormones covalently bound to glass and Sepharose beads.
Isoproterenol, corticotropin (ACTH), and triodothyronine immobilized on glass and Sepharose beads by diazotization procedures have been shown to interact with cultured tumor cells of "target tissue" origin. Cells used were rat glioma cells (C6), rat adrenal tumor cells (Y-1), and rat pituitary tumor cells (GH3). The rat glioma cells bound principally to immobilized isoproterenol, whereas the rat adrenal tumor cells bound to immobilized corticotropin, and rat pituitary tumor cells bound to immobilized triiodothyronine. Binding was inhibited by preincubation of the cells in soluble drug or hormone. With C6 cells there was a positive correlation between adenylate cyclase [ATP pyrophosphate-lyase (cyclizing, EC 4.6.1.1] stimulation and the degree of binding to the immobilized isoproterenol. Norepinephrine, bound through the ethanolamine side chain via an amide linkage, did not bind cells, demonstrating specific structural requirements for drug-cell interactions. HeLa cells were shown to bind tightly to diphtheria toxin coupled to Sepharose beads via an amide bond. This binding was inhibited by prior incubation of the Sepharose toxin with purified antitoxin. Toxin bound to Sepharose via an azo bond did not bind cells. These data suggest that the cell affinities are due to cell surface receptors interacting with the immobilized drugs and hormones, and that the observed affinities possibly reflect the relative receptor complement of these cells.
D000310:Adrenal Gland Neoplasms / Q000378:metabolism; D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods; D004167:Diphtheria Toxin / Q000378:metabolism; D005910:Glioma / Q000378:metabolism; D006367:HeLa Cells; D007545:Isoproterenol / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000378:metabolism* / Q000473:pathology; D010911:Pituitary Neoplasms / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; D014284:Triiodothyronine / Q000378:metabolism
D004167:Diphtheria Toxin; D011956:Receptors, Cell Surface; D011955:Receptors, Drug; D014284:Triiodothyronine; D000324:Adrenocorticotropic Hormone; D007545:Isoproterenol