Immunological and physiological analysis of human breast cancer progesterone receptor heterogeneity, following KCl dissociation and size exclusion chromatography.
Progesterone receptor (PR) levels were determined in 69 human breast cancer specimens by both radioligand assay (RLA) and enzyme immunoassay (EIA). These methods did not detect the same number of sites, and for each tumor there was a constant ratio between epitopes and PR-binding sites corresponding to 1/4, 2/4, ... 8/4. High performance size exclusion chromatography was performed to separate the various PR isoforms, and the ability of these isoforms to interact with the monoclonal antibodies was assessed. Determination of PR in the chromatographic fractions by EIA and RLA showed that the various isoforms isolated by chromatography presented variable quantities of steroid-binding sites and epitopes, thus confirming the differences observed in the cytosol assays. The dissociation of molybdate-stabilized PR by KCl and measurement by RLA and EIA of the isoforms obtained showed two different types of chromatographic patterns, particularly in the 8S polymeric form where the monoclonal antibodies appeared to detect mainly the heavier 8S fraction, which may correspond to the 8S-B form of the progesterone receptor. The monoclonal antibodies also detected an intermediate PR polymeric form (236 kDa) which was not always detected by the tritiated ligand. Our results suggest that breast cancer PR exhibit a marked molecular heterogeneity which may partially explain the differences in response to hormonal therapy, particularly for tumors with high receptor levels which nonetheless fail to respond to treatment.
Goussard
J
J
Laboratoire d'Analyses Isotopiques, Centre François Baclesse, Caen, France.
Lechevrel
C
C
Roussel
G
G
D000911:Antibodies, Monoclonal; D001665:Binding Sites; D001943:Breast Neoplasms / Q000378:metabolism*; D002850:Chromatography, Gel; D046508:Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007202:Indicators and Reagents; D011189:Potassium Chloride; D011863:Radioimmunoassay; D011980:Receptors, Progesterone / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007202:Indicators and Reagents; D011980:Receptors, Progesterone; D011189:Potassium Chloride