Characterization of a receptor-negative, hormone-nonresponsive clone derived from a T47D human breast cancer cell line kept under estrogen-free conditions.
We have established an estrogen receptor- and progesterone receptor-negative, hormone-nonresponsive breast cancer cell line from a receptor-positive, hormone-responsive line grown under estrogen-free conditions. T47D breast cancer cells were cultured under estrogenized conditions (in phenol red-containing medium supplemented with whole fetal bovine serum) and cloned to produce line T47D:A18. The parental T47D line was also estrogen deprived (in phenol red-free medium supplemented with dextran-coated charcoal-treated fetal bovine serum) for more than 1 year and subsequently clone T47D:C4 was established. T47D:A18 was estrogen receptor and progesterone receptor positive as determined by both ligand binding assay analysis and enzyme immunoassay analysis. T47D:C4 cells were estrogen receptor and progesterone receptor negative and mRNA for these receptors was not detected. Incubation of hormone-responsive T47D:A18 cells with 17 beta-estradiol caused a 3-fold increase in cell growth over 8 days when compared to control. This stimulation of growth was completely inhibited by the anti-estrogens 4-hydroxytamoxifen (0.1 microM) and ICI 164,384 (1.0 microM). Receptor-negative T47D:C4 cells were refractory to the effects of both 17 beta-estradiol and the antiestrogens. T47D:A18 cells grown under both estrogen-containing and estrogen-free conditions expressed low levels of transforming growth factor (TGF)-alpha and epidermal growth factor receptor mRNA. In the presence of estrogen, high levels of TGF-beta 1 mRNA were detected in T47D:A18 cells. These levels decreased when T47D:A18 cells were grown in estrogen-free media. Conversely, TGF-beta 2 mRNA was not detected in T47D:A18 cells cultured under estrogenic conditions; however, message was detected after the cells were cultured under estrogen-free conditions. T47D:C4 cells expressed low levels of TGF-alpha, epidermal growth factor receptor, TGF-beta 1, and TGF-beta 2 mRNA. These studies characterize a novel hormone-nonresponsive cell line which has been established from a hormone-responsive cell line grown under estrogen-free and drug-free conditions. Further analysis of these lines should provide valuable information concerning the development of antiestrogen-resistant breast cancer.
Murphy
C S
CS
Department of Human Oncology, University of Wisconsin, Madison 53792.
Pink
J J
JJ
Jordan
V C
VC
D015152:Blotting, Northern; D001943:Breast Neoplasms / Q000473:pathology*; D002455:Cell Division / Q000187:drug effects; D002999:Clone Cells; D003562:Cytidine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000633:toxicity; D016172:DNA Fingerprinting; D004273:DNA, Neoplasm / Q000032:analysis; D066246:ErbB Receptors / Q000235:genetics; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics / Q000502:physiology*; D011980:Receptors, Progesterone / Q000235:genetics; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D016211:Transforming Growth Factor alpha / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D053284:Polyunsaturated Alkamides; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D016211:Transforming Growth Factor alpha; D016212:Transforming Growth Factor beta; D013629:Tamoxifen; C016601:afimoxifene; D004958:Estradiol; D003562:Cytidine; C045911:cyclopentenyl cytosine; C051781:ICI 164384; D066246:ErbB Receptors