Multiple isoforms of the oestrogen receptor in endometrial cancer.
We evaluated the presence and variability of oestrogen receptor (ER) isoforms in endometrial cancer by using [3H]oestradiol-labelled ERs and the H222 monoclonal antibody obtained from the Abbott enzyme immunoassay kit. Using isoelectric focusing (IEF), endometrial ER was shown to be composed of four different species, with pI values of 6.1, 6.3, 6.6 and 6.8, indistinguishable from the isoforms found in normal rat uterus, and human breast and larynx carcinomas. The isoforms at pI 6.3, 6.6 and 6.8, all sedimenting at 4S by sucrose gradient fractionation, showed, on two-dimensional SDS electrophoresis, relative masses of 50, 70 and 65 kDa respectively, equal to the masses previously found in breast cancer. These isoforms did not alter their pI values during IEF fractionation performed in a linear gradient of urea, while the pI 6.1, sedimenting at 8S, generated a new isoform at about 9 mol/l urea with pI 7.2 and a relative mass of 65 kDa. The urea-dissociated isoform (pI 7.2) was able approximately to double the antibody binding with respect to the nondissociated oligomer, which suggested that some epitopes are 'masked', i.e. not accessible to the antibodies when ER is present in its complexed form. The evidence thus suggested that the oligomer at pI 6.1 contained a single 65 kDa ER form which, as a monomer, focused at pI 7.2. The variability in the ER isoform profile found in endometrial cancer was similar to the variability previously reported in breast and larynx carcinomas. The balance between these isoforms could be a dynamic parameter involved in the functionality of this receptor and consequently in cell transformation.
Marsigliante
S
S
Dipartimento di Biologia, Università di Lecce, Via Prov. le per Monteroni, Italy.
Muscella
A
A
Ciardo
V
V
Puddefoot
J R
JR
Leo
G
G
Vinson
G P
GP
Storelli
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000737:chemistry; D002277:Carcinoma / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D016889:Endometrial Neoplasms / Q000737:chemistry*; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007822:Laryngeal Neoplasms / Q000737:chemistry; D009363:Neoplasm Proteins / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D051381:Rats; D011960:Receptors, Estrogen / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D014599:Uterus / Q000737:chemistry
D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; D011960:Receptors, Estrogen