Comparison of histochemical and biochemical assays for estrogen receptor in human breast cancer cell lines

Two human breast cancer lines, MCF-7 and T47D cells, were investigated for the presence of estrogen receptor (ER) by biochemical and histochemical techniques. Using the dextran-coated charcoal technique and isoelectric focusing, MCF-7 cells were ER positive, and T47D cells were ER negative. Fluoresc...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1984, Vol.44 (1), p.415-421
Main Authors: PARL, F. F, WETHERALL, N. T, HALTER, S, SCHUFFMAN, S, MITCHELL, W. M
Format: Article
Language:English
Subjects:
Biological and medical sciences
Breast Neoplasms - metabolism
Breast Neoplasms - pathology
Cell Line
Estradiol - metabolism
Female
Fluorescent Dyes
Gynecology. Andrology. Obstetrics
Humans
Isoelectric Focusing
Mammary gland diseases
Medical sciences
Receptors, Estrogen - isolation & purification
Receptors, Estrogen - metabolism
Tritium
Tumors
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Summary:Two human breast cancer lines, MCF-7 and T47D cells, were investigated for the presence of estrogen receptor (ER) by biochemical and histochemical techniques. Using the dextran-coated charcoal technique and isoelectric focusing, MCF-7 cells were ER positive, and T47D cells were ER negative. Fluorescein conjugates to 17 beta-estradiol by the sixth carbon (17 beta-estradiol-6-carboxymethyloxime:bovine serum albumin: fluorescein isothiocyanate and 17 beta-estradiol-6-iminooxyacetylfluoresceinamine) and by the 17th carbon [N-fluoresceino-N'-[17 beta-(estradiol hemisuccinamide)ethyl]thiourea, 17-FE] were prepared for cytochemical evaluation of the ER status of the two cell lines. The binding affinity of the estradiol conjugates for ER varied, the 17-FE conjugate having the highest affinity of 0.08 relative to 17 beta-estradiol. Following incubation with 10 nM 17-FE, both MCF-7 and T47D cells displayed cytoplasmic and nuclear fluorescent staining. Isoelectric focusing of MCF-7 cytosol incubated in the presence of 10 nM 17-FE revealed binding of the fluorescein conjugate to a protein species which did not bind 17 beta-[3H]-estradiol. Isoelectric focusing of T47D cytosol revealed binding of 17-FE to two protein components, neither one of which showed specific binding of 17 beta-[3H]estradiol. The results suggest different protein binding species for fluoresceinated estradiol conjugates and [3H]estradiol and help to explain reported differences in histochemical and biochemical ER analyses.
ISSN:0008-5472
1538-7445