HPLC-RIA analysis of steroid hormone profile in a virilizing stromal tumor of the ovary.
The pathological steroid biosynthesis of a virilizing ovarian tumor was examined via high performance liquid chromatography-radioimmunoassay (HPLC-RIA) determination of the intratissular steroid concentrations. Sex cord-stromal tumor of the ovary was obtained surgically from an 18-year-old female patient with extremely high androst-4-ene-3,17-dione (4-en-dione) and testosterone (Test) blood serum levels. The tissue specimen was extracted with ethyl acetate and the extract was then purified on a C18 mini-column with methanol-water eluents. Steroids were isolated by reversed-phase HPLC on a C18 silica gel column with 51%, 55% and 64% v/v methanol-water eluents. Steroids in the collected eluent fractions were detected by the radioactivity of tritiated internal standards and then quantified by specific RIAs. In the tumor specimen, very high 17alpha-hydroxyprogesterone (17-OH-Prog; 6300 fmol/g), dehydro-epiandrosterone (2870 fmol/g), androst-4-ene-3,17-dione (3000 fmol/g), testosterone (5700 fmol/g) concentrations, and less progesterone (PROG; 320 fmol/g) and androst-5-ene-3beta,17beta-diol (5-en-diol; 320 fmol/g), were determined. Tissue levels of 5alpha-dihydrotestosterone (DHT), 5alpha-androstane-3alpha,17beta-diol (3alpha-diol), 5alpha-androstane-3beta,17beta-diol (3beta-diol), and 17beta-estradiol were found to be 71, 20, 28, and 12 fmol/g, respectively. Steroid profile analysis verified a pathological steroid biosynthesis in the ovarian tumor and suggested that the 17alpha-hydroxylase (17alpha-H), 17,20-lyase (17,20-L), and 3beta-hydroxysteroid dehydrogenase/Delta5-4-isomerase (Delta5-3beta-HSD) activities were particularly elevated in this tumorous tissue. Present data demonstrate that the analysis of intratissular steroid profile by a HPLC-RIA method may valuably contribute to the steroidal pathophysiology of endocrine tumors.
Szécsi
M
M
Endocrine Unit and Research Laboratory, University of Szeged, 8 Korányi fasor, H-6720 Szeged, Hungary. szecsim@endoc.szote.u-szeged.hu
Tóth
I
I
Gardi
J
J
Nyári
T
T
Julesz
J
J
D000293:Adolescent; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D036821:Endometrial Stromal Tumors / Q000150:complications / Q000378:metabolism*; D005260:Female; D012739:Gonadal Steroid Hormones / Q000032:analysis* / Q000378:metabolism*; D006801:Humans; D010051:Ovarian Neoplasms / Q000150:complications / Q000378:metabolism*; D011863:Radioimmunoassay / Q000379:methods*; D014407:Tumor Cells, Cultured; D014770:Virilism / Q000209:etiology / Q000378:metabolism*
D014408:Biomarkers, Tumor; D012739:Gonadal Steroid Hormones