The effect of 1.25 dihydroxyvitamin D3 on binding and internalization of epidermal growth factor in cultures cells. Studies on BT-20 cells using quantitative electron microscope autoradiography.
The biological effects of 1.25 (OH)2D3 on epidermal growth receptor (EGF-R) and on EGF internalization were examined in human mammary carcinoma BT-20 cells. In this cell line, with known amplification of the epidermal growth factor receptor gene. EGF was not stimulatory for growth. Biological assay and quantitative EM autoradiography combined with iodinated ligand binding to specific receptors demonstrated that the number of binding sites unit of length of plasma membrane was 2.48-fold higher in treated than in control cells. I-EGF was progressively internalized in a time-and temperature-dependent manner after selective association with the membrane-coated pits. No modification of the time course of I-EGF internalization was noted in the control and in the treated cells, but a different distribution of the labeling in the subcellular compartment was observed in treated cells. In 1.25(OH)2D3-treated batches, the grain density remained low in the receptosomes throughout the experiment, whereas it was high and occurred early in the lysosomes. On the other hand, in control cells, the grain density of the receptosomes was high, whereas it occurred late and was relatively low in the lysosomes. These data suggest that 1.25(OH)2D3 is a regulator of EGF-R level in BT-20 cell line, but it cannot affirmed whether this effect is direct or mediated by other parameters.
Frappart
L
L
Laboratoire de Biologie Médicale, Centre Régional de Lutte contre le Cancer, Lyon, France.
Lefebvre
M F
MF
Saez
S
S
D001345:Autoradiography / Q000379:methods; D001940:Breast / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D002117:Calcitriol / Q000494:pharmacology* / Q000502:physiology; D002277:Carcinoma / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D002455:Cell Division; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D007457:Iodine Radioisotopes / Q000378:metabolism; D008854:Microscopy, Electron / Q000379:methods; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure
D007457:Iodine Radioisotopes; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D002117:Calcitriol