Characterization and assay of progesterone-binding components in DMBA-induced rat mammary carcinoma tissue after progesterone administration

Abstract

Nuclear and cytoplasmic progesterone-binding components were characterized and measured in DMBA-induced rat mammary carcinoma tissue, before and after progesterone administration. Rats, bearing growing tumors, were ovariectomized and then primed for two days with estradiol. Biopsy specimens were taken prior to or following administration of progesterone. Nuclear binding was assayed in the 0.4 M KCl extract of the nuclear fraction using [3H]R5020 as ligand. The receptor character of the binding was demonstrated by: (1) high affinity (Kd approximately 2 nM); (2) specificity: competition by R5020 and progesterone, minimal competition by 17 alpha-hydroxyprogesterone, corticosterone, testosterone and estradiol; (3) sedimentation constant at about 3S in a sucrose density gradient. Similar characteristics displayed the cytoplasmic receptor before and after progesterone administration. Progesterone receptor distribution in the nuclear extract and cytosol were determined in 36 tumors. The levels of total receptors (cytoplasmic plus nuclear) before and after progesterone administration varied widely, however the average values found after progesterone administration were significantly lower, 1.59 +/- 0.20 pmol/mg DNA compared to 2.58 +/- 0.32 pmol/mg DNA. Before progesterone administration only cytoplasmic receptors were found. One hour after progesterone administration a variable amount of the receptor (0-40%) was found in the nucleus of the tumorous tissue. In uteri of the same rats a uniform distribution of receptors (about 40% in the nucleus) was found after progesterone administration. A defect in the translocation process might be considered in the tumors with low receptors level, which suggests that DMBA-tumors may not respond uniformly to progesterone administration.