Comparison of the in vitro conversion of estradiol-17 beta to estrone of normal and neoplastic human breast tissue.
Specific activity  of 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) was measured in 48 tissue specimens of human female breast cancer and, in addition, 48 nonmalignant tissue specimens obtained in each case from the same cancer-bearing breast. In all cases the nonmalignant tissue showed greater conversion of estradiol-17 beta into estrone than the neoplastic tissues. In normal human breast tissue of premenopausal women specific enzyme activity depended on the phase of the MENSTRUAL CYCLE: the highest values of 17 beta-HSD activity were found in the early secretory phase. To determine the intracellular distribution of the 17 beta-HSD, purified microsomes, mitochondria, peroxysomes, lysosomes, nuclei and cytosol fractions were prepared. The purity of each fraction was monitored by marker enzymes. It was found that the 17 beta-HSD was mainly located in mitochondria and microsomes. Furthermore it could be demonstrated that the microsomal enzyme was bound tightly to the membranes of the endoplasmic reticulum, while the mitochondrial 17 beta-HSD was mainly associated with the outer membranes of the organelle. Kinetic parameters (Km-values, coenzyme requirements and maximal velocities) of a cytoplasmic, nuclear, mitochondrial and microsomal 17 beta-HSD of normal and neoplastic human mammary tissue were compared. Maximal velocity was highest in enzyme preparations of normal mammary tissue obtained from premenopausal women in the early secretory phase. Km-values wrere nearly identical in normal and neoplastic mammary tissue preparations (approx. 1 X 10(-6) M). NAD was more efficient than NADP as a cofactor. For the conversion of estradiol to estrone the optimum temperature was approximately 40 degrees C and the optimum pH 9.5. For the reduction of estrone the optimum pH was 6.5. Sulphydryl groups were shown to be essential for catalysis.
D001940:Breast / Q000201:enzymology* / Q000648:ultrastructure; D001943:Breast Neoplasms / Q000201:enzymology*; D004721:Endoplasmic Reticulum / Q000201:enzymology; D004958:Estradiol / Q000378:metabolism; D004960:Estradiol Dehydrogenases / Q000378:metabolism*; D004970:Estrone / Q000378:metabolism; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006913:Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D066298:In Vitro Techniques; D007700:Kinetics; D008593:Menopause; D008598:Menstruation; D008861:Microsomes / Q000201:enzymology; D008928:Mitochondria / Q000201:enzymology; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013696:Temperature
D013439:Sulfhydryl Reagents; D004970:Estrone; D004958:Estradiol; D006913:Hydroxysteroid Dehydrogenases; D004960:Estradiol Dehydrogenases