Antiestrogen stimulation of the production of a 37,000 molecular weight secreted protein and estrogen stimulation of the production of a 32,000 molecular weight secreted protein in MCF-7 human breast cancer cells.
Antiestrogens (AE), such as tamoxifen, selectively increase the production of a secreted protein of 37,000 mol wt (Mr) in estrogen receptor-containing human breast cancer cells (MCF-7, but not in estrogen receptor-negative MDA-MB-231 cells), and the production of this protein by AE is inhibited by concomitant estradiol (E) treatment. Likewise, E increases the production of a 32,000 Mr secreted protein whose synthesis is inhibited by AE. Proteins were detected by [35S]methionine and [35S]cysteine labeling of cells and analysis of proteins by sodium dodecyl sulfate-polyacrylamide gels and fluorography. Enhanced production of the 37,000 Mr protein is observed within 6 h of AE treatment, with maximal synthesis seen at 1-2 days when this protein represents about 6% of the total radiolabeled secreted proteins. This protein is stimulated maximally (approximately 4-fold) by 10(-8) M trans-hydroxytamoxifen or LY117018 or 10(-6) M tamoxifen, and its AE specificity is seen by the fact that trans-tamoxifen increases this protein, whereas cis-tamoxifen, an estrogen, does not. In addition to stimulating the synthesis of previously identified 160,000 and 52,000 Mr secreted proteins, E increases the production of a 32,000 Mr secreted protein. When cells are grown in estrogen-free conditions, i.e. in charcoal-dextran-treated serum in medium lacking the estrogen phenol red, the basal level of the 32,000 Mr protein is extremely low, and E stimulation results in a 10-fold increase in the production of this protein, with increases observable by 6 h and maximal stimulation at 2 days. Interestingly, the basal level of synthesis of the 37,000 Mr protein is high in the absence of E and is then stimulated only minimally by the addition of AE, suggesting that this protein is clearly produced as an estrogen-antagonistic protein. Amino acid incorporation conducted in the presence of tunicamycin and endoglycosidase H indicates that both of these proteins are glycoproteins. These proteins should serve as useful markers for AE and E action and may be involved in AE and E modulation of cell proliferation and/or cell function.
D000596:Amino Acids / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism*; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D006801:Humans; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D013462:Sulfur Radioisotopes; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology*
D000596:Amino Acids; D009363:Neoplasm Proteins; D013462:Sulfur Radioisotopes; D013629:Tamoxifen; D004958:Estradiol; D008715:Methionine; D003545:Cysteine