Microassay for nuclear binding of steroid receptors with use of intact cells from small samples of avian and human tissue.
A micro version of a nuclear binding assay to assess the biological activity of receptors for steroid hormones was developed for application to small (needle) biopsies of human tumors for the purpose of predicting responses to steroid therapy. This easier assay requires 10-fold less tissue than the original nuclear binding assay described for progesterone receptors in the avian oviduct, endometrium, and endometrial carcinomas (Spelsberg TC, et al., Endocrinology 1987;121:631). We describe the application of this micro assay to normal avian oviduct and cancers of the human breast, and we demonstrate a tissue specificity and saturation of nuclear binding. The micro assay reliably measured as little as 0.5 mg equivalents of tissue per assay tube. Results for breast tumors determined to be estrogen-receptor-positive by the standard dextran-coated charcoal method were also determined with this nuclear binding assay. As described previously for progesterone receptors in endometrial carcinomas, some receptor-positive breast biopsies displayed negligible capacity for nuclear binding. Therefore, with the present assay we have identified nonfunctional receptors in these biopsies, which may be useful for accurate prediction of patients' responses to therapy with hormones.
Colvard
D S
DS
Department of Biochemistry and Molecular Biology, Mayo Graduate School of Medicine, Mayo Clinic, Rochester, MN 55905.
Jankus
W R
WR
Berg
N J
NJ
Graham
M L
ML
Jiang
N S
NS
Ingle
J N
JN
Spelsberg
T C
TC
D000818:Animals; D001667:Binding, Competitive; D001707:Biopsy, Needle; D001943:Breast Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D002645:Chickens; D004247:DNA / Q000032:analysis; D004958:Estradiol / Q000378:metabolism; D006728:Hormones / Q000627:therapeutic use; D006801:Humans; D008832:Microchemistry; D010057:Oviducts / Q000378:metabolism*; D011397:Promegestone / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D011980:Receptors, Progesterone / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism*
D006728:Hormones; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D011987:Receptors, Steroid; D004958:Estradiol; D004247:DNA; D011397:Promegestone