Summary
Rabbit antibodies to cytoplasmic estrogen receptors (ER) of human breast carcinoma were utilized for investigating steroid-triggered in-vitro translocation of cytoplasmic ER to the nuclear compartment of the estrogen target cells. The immunofluorescent method (IF) previously described (S Raam et al., Eur J Cancer Clin Oncol 18: 1–12, 1982) was employed for immunohistochemical localization of ER. Four cases of normal endometrium, two cancers of the endometrium, and MCF-7 human breast cancer cells were maintained in a steroid free medium and exposed at 37°C for two hours to growth medium alone (control) or to 2.5, 25 or 250 nanomoles of estradiol (E2), diethylstilbestrol (DES), or monohydroxytamoxifen (OH-TX). At the end of the incubation period the cells were processed for intracellular localization of ER.
Complete traslocation of IF from the cytoplasm to the nuclear compartment was evident in all normal endometrial cells exposed to E2, DES or OH-TX for two hours. While cells from the endometrial cancer ‘S’, like the normal cells, translocated IF to the nucleus, cells of another cancer (‘KLE’) failed to translocate when exposed to E2 or OH-TX. Partial translocation was evident in ‘KLE’ cells exposed to DES.
In MCF-7 cells grown in the absence of E2, IF was exclusively cytoplasmic. When these cells were exposed to the hormones, 50% showed a complete transfer of IF to the nucleus; in 40% a delayed response was evident; 10% failed to translocate.
The results revealed the suitability of anti-ER antibodies for investigating the intracellular dynamics of ER in target cells responding to estrogens or antiestrogens.
References
Muldoon TG: Regulation of steroid hormone receptor activity. Endocr Rev 1: 339–364, 1980
Mercer WD, Lippman ME, Wahl TM, Carlson CA, Wahl DA, Lezotte D, Teague PO: The use of immunocytochemical techniques for the detection of steroid hormones in breast cancer cells. Cancer 46(Suppl.): 2859–2868, 1980
Raam S, Peters L, Forcier A, Cohen JL: Antibodies to human breast tumor estrophilin. Poster presented at the Symposium on Steroid Hormone Receptor Systems, Worcester Foundation for Experimental Biology 1979.
Raam S, Peters, L, Rafkind I, Putnam E, Longcope C, Cohen JL: Simple methods for production and characterization of rabbit antibodies to human breast tumor estrogen receptors. Mol Immunol 18: 143–156, 1981
Senior MB, Raam S, Sun L, Frankel FR: Studies on the interaction between estradiol bearing chromatin components from MCF-7 cells and a preparation of estradiol receptor antibodies. Biochemistry (submitted)
Raam S, Nemeth EJ, Tamura H, O'Briain DS, Cohen JL: Immunohistochemical localization of estrogen receptors in human mammary carcinoma using antibodies to the receptor protein. Eur J Cancer Clin Oncol 18: 1–12, 1982
Satyaswaroop PG, Bressler RS, DeLaPena MM, Gurpide E: Isolation and culture of human endometrial glands. J Clin Endocrinol Metab 48: 639–641, 1978
Kelner KL, Miller AL, Peck EJ: Estrogen and hypothalamus: Nuclear receptor and RNA polymerase activation. J Receptor Res 1: 215–237, 1980
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin-phenol reagent. J Biol Chem 193: 265–275, 1951
Nenci I, Beccati MD, Piffanelli A, Lanza G: Detection and dynamic localisation of estradiol-receptor complexes in intact target cells by immunofluorescence technique. J Steroid Biochem 7: 505–510, 1976
Chamness GC, Mercer WD, McGuire WL: Are histochemical methods for estrogen receptor valid? J Histochem Cytochem 28: 792–798, 1980
Eckert RL, Katzenellenbogen BS: Effects of estrogens and antiestrogens on estrogen receptor dynamics and the induction of progesterone receptor in MCF-7 human breast cancer cells. Cancer Res 42: 139–144, 1982
Nawata H, Bronzert D, Lippman ME: Isolation and characterization of a tamoxifen-resistant cell line derived from MCF-7 human breast cancer cells. J Biol Chem 256: 5016–5021, 1981
Author information
Authors and Affiliations
Additional information
Address for reprints: Shanthi Raam, Ph.D., Tufts University Medical Cancer Unit, Lemuel Shattuck Hospital, 170 Morton Street, Boston, MA 02130.
Rights and permissions
About this article
Cite this article
Raam, S., Richardson, G.S., Bradley, F. et al. Translocation of cytoplasmic estrogen receptors to the nucleus: Immunohistochemical demonstration utilizing rabbit antibodies to estrogen receptors of mammary carcinomas. Breast Cancer Res Tr 3, 179–199 (1983). https://doi.org/10.1007/BF01803561
Issue Date:
DOI: https://doi.org/10.1007/BF01803561