Free and Conjugated Estrogens of an Ovarian Cyst adenoma and Granulosa Cell Tumor

Abstract

A granulosa cell tumor, from a patient with signs of hyperestrinism, was incubated 4 hr in Krebs-Ringer solution. Free steroids, glucoronosides and sulfates were sequentially hydrolzed with enzymes and extracted. Cyst fluid (200 ml) from a mucinous cystadenoma was similarly treated. Phenols were fractionated initially on alumina columns. Zones on paper chromatograms which paralleled reference estrone and estradiol-17[beta], and reacted with potassium ferricyanide-feric chloride, were eluted and the Kober reaction was performed. Gas chromatography further supported the identity of the estrogens and was used for quantitation. The granulosa tumor incubate contained 0.25 and 0.1 [mu]g/g/hr of free estradiol-17[beta] and estrone, respectively. Estrogens were not detected following hydrolysis with [beta]-glucuronidase. The concentration of estron following hydrolysis with phenol sulfatase was 1.0 [mu]g/g/hr. Estradiol-17[beta] and estriol were not present in the sulfate fraction. Estrogens were not found in the free or glucuronoside fractions of the cyst fluid. The estrone fulfate concentration was 10 [mu]g/100 ml. The high concentration of estrone sulfate in these tumors indicates that this conjugate may play a role in the metabolism of estrogens by ovarian tissue.