Effects of TCDD on the EGF receptor of XB mouse keratinizing epithelial cells.
TCDD was found to cause a marked inhibition of 125I-epidermal growth factor (EGF) binding to its receptor on the cell surface of XB mouse keratinizing epithelial cells (XB cells) cultured in vitro. The EC50 concentration was estimated to be on the order of 3 x 10(-11) M 24 hours after TCDD administration. As early as 12 hours after the addition of 10(-9) M of TCDD, XB cells showed signs of a decline in 125I-EGF binding levels. The level of such EGF receptor downregulation reached a maximum at 24 hours, continued until day 2, but completely recovered by day 3. This was accompanied by a rise in protein kinase activities, particularly those of the protein tyrosine kinases during the initial period of 6-24 hours. To test the hypothesis that the EGF receptors of the cells, by showing TCDD-induced symptoms of downregulation, actually are being activated and triggering EGF-like signals, we examined the effects of both TCDD and exogenously added EGF on cell morphology, colony formation degree of keratinization, the pattern of activation of protein kinases and de novo protein synthesis, and EGF receptor phosphorylation. Based on the similarity of cell responses to these between TCDD- and EGF-treated cells, we concluded that TCDD, directly or indirectly, causes activation of the EGF receptor. In contrast, 12-O-tetradencanoylphorbol-13-acetate (TPA), which is known to downregulate EGF receptors by blocking their protein tyrosine kinase, produced dissimilar end results. The balance of evidence support the notion that the action of TCDD in this cell line is tightly coupled to the activation of the EGF receptor and that one of the key consequences of such a biochemical change is that it signals these cells to commit to terminal differentiation.
Kawamoto
T
T
Department of Environmental Toxicology, University of California, Davis 95616.
Matsumura
F
F
Madhukar
B V
BV
Bombick
D W
DW
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004147:Dioxins / Q000633:toxicity*; D015536:Down-Regulation / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D066246:ErbB Receptors / Q000187:drug effects*; D007457:Iodine Radioisotopes; D008715:Methionine / Q000378:metabolism; D051379:Mice; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity*; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D013194:Staining and Labeling; D013462:Sulfur Radioisotopes
D004147:Dioxins; D007457:Iodine Radioisotopes; D000072317:Polychlorinated Dibenzodioxins; D013462:Sulfur Radioisotopes; D008715:Methionine; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases