Tyrphostins Inhibit Epidermal Growth Factor (EGF)-Receptor Tyrosine Kinase Activity in Living Cells and EGF-Stimulated Cell Proliferation

• Published in: The Journal of biological chemistry 1989-08, Vol.264 (24), p.14503-14509
• Main Authors: Lyall, R M, Zilberstein, A, Gazit, A, Gilon, C, Levitzki, A, Schlessinger, J
• Format: Article
• Language: English
• Subjects: 3T3 cells; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Catechols - pharmacology; Cell Division - drug effects; Cell Line; DNA - biosynthesis; Endocytosis - drug effects; Enzymes and enzyme inhibitors; Epidermal Growth Factor - metabolism; Epidermal Growth Factor - pharmacology; ErbB Receptors - drug effects; ErbB Receptors - metabolism; Fibroblasts - enzymology; Fibroblasts - metabolism; Fibroblasts - physiology; Fundamental and applied biological sciences. Psychology; Growth Inhibitors - pharmacology; Kinetics; Mice; mitogenicity; Nitriles - pharmacology; Phosphorylation; Protein-Tyrosine Kinases - antagonists & inhibitors; Transferases; Tyrosine - analogs & derivatives; Tyrosine - pharmacology; Tyrphostins
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)71707-9

Synthetic compounds called tyrphostins were examined for their effects on cells which are mitogenically responsive to epidermal growth factor (EGF). We studied in detail the effects of two tyrphostins on EGF binding, tyrosine phosphorylation in intact cells, EGF-receptor internalization, and mitogenesis. These compounds inhibited EGF-stimulated [3H]thymidine incorporation in a specific manner and the degree of selectivity varied. Both compounds inhibited EGF-stimulated receptor autophosphorylation and tyrosine phosphorylation of endogenous substrates in intact cells at doses that correlated with the IC50 for [3H] thymidine incorporation. These results are consistent with the notion that tyrosine phosphorylation is a crucial signal in transduction of the mitogenic message delivered by EGF. The compound RG50864 demonstrated specificity at inhibiting EGF-stimulated cell growth compared with stimulation with either platelet-derived growth factor or serum. For both compounds RG50864 and RG50810, long term exposure (16 h) of cells to tyrphostins was required for optimal inhibition because of the instability and slow action of these compounds. Tyrphostins did not alter cell surface display of EGF-receptor, EGF binding or EGF-induced internalization, degradation, and down-regulation of EGF receptors. These novel synthetic inhibitors, specific for EGF-receptor kinase, offer a new method to inhibit EGF-stimulated cell proliferation which may be useful in treating specific pathological conditions involving cellular proliferation, including different types of cancers.