TY - JOUR
T1 - Altered phenotypic characteristics of T47D human breast cancer cells after prolonged growth in estrogen-deficient medium
AU - Fernandez, Pat
AU - Wilson, Cody
AU - Hoivik, Debie
AU - Safe, Stephen H.
PY - 1998/9
Y1 - 1998/9
N2 - T47D human breast cancer cells were cultured in estrogen-deficient media for up to 32 months and the resulting cell line (L(hE-)) exhibited unique phenotypic and genotypic characteristics. Compared to low passage (L) cells, the L(hE-) cells exhibited a significantly higher rate of proliferation, unique morphological features, advanced ploidy status and 5- to 10-fold higher levels of the estrogen receptor (ER) as determined by ligand binding and Western blot analysis. Sequence analysis of the DNA binding domain of the ER revealed a C→A transversion which resulted in a H513N amino acid change. Treatment of L cells with 10 nM 17β-estradiol (E2) resulted in a greater than two-fold increase in cell proliferation which was inhibited by tamoxifen, 4'-hydroxytamoxifen, ICI 164,384 and ICI 182,780. In contrast, 10 nM E2 caused a 70% decrease in growth of L(hE-) cells and this antimitogenic activity was blocked by ICI 164,384 and ICI 182,780 but not by tamoxifen or 4'-hydroxytamoxifen. L(hE-) cells were E2-responsive in transient transfection studies using a plasmid containing an estrogen-responsive element derived from the vitellogenin A2 gene promoter. These data show that the phenotypic and genotypic characteristics of L(hE-) T47D cells resemble those described for ER-negative cell lines stably transfected with the ER.
AB - T47D human breast cancer cells were cultured in estrogen-deficient media for up to 32 months and the resulting cell line (L(hE-)) exhibited unique phenotypic and genotypic characteristics. Compared to low passage (L) cells, the L(hE-) cells exhibited a significantly higher rate of proliferation, unique morphological features, advanced ploidy status and 5- to 10-fold higher levels of the estrogen receptor (ER) as determined by ligand binding and Western blot analysis. Sequence analysis of the DNA binding domain of the ER revealed a C→A transversion which resulted in a H513N amino acid change. Treatment of L cells with 10 nM 17β-estradiol (E2) resulted in a greater than two-fold increase in cell proliferation which was inhibited by tamoxifen, 4'-hydroxytamoxifen, ICI 164,384 and ICI 182,780. In contrast, 10 nM E2 caused a 70% decrease in growth of L(hE-) cells and this antimitogenic activity was blocked by ICI 164,384 and ICI 182,780 but not by tamoxifen or 4'-hydroxytamoxifen. L(hE-) cells were E2-responsive in transient transfection studies using a plasmid containing an estrogen-responsive element derived from the vitellogenin A2 gene promoter. These data show that the phenotypic and genotypic characteristics of L(hE-) T47D cells resemble those described for ER-negative cell lines stably transfected with the ER.
KW - Estrogen growth-inhibition
KW - T47D variant
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U2 - 10.1006/cbir.1998.0303
DO - 10.1006/cbir.1998.0303
M3 - Article
C2 - 10452832
AN - SCOPUS:0032157953
VL - 22
SP - 623
EP - 633
JO - Cell Biology International
JF - Cell Biology International
SN - 1065-6995
IS - 9-10
ER -