Testosterone metabolism in the human endometrium : a combination of metabolic (mass spectrometry) and enzyme expression (RT-PCR)

Localised steroid metabolism is important in proliferative disorders of the endometrium. The metabolism of testosterone, an important precursor in endometrial disorders, was investigated by mass spectrometry and RT-PCR (real-time polymerase chain reaction), allowing determination of steroid metabolites and expression of steroid converting enzymes; aromatase, 17P-HSD1, 2,4,5,7,8,(hydroxysteroid dehydrogenase) and 5AR1, 2 (alpha reductase). Samples were derived from cell lines (Ishikawa, HEC-1A, HEC-1B, RL95-2, COV434), and biopsies; fertile, endometriosis, poly-cystic ovary syndrome (PCOS), endometrial hyperplasia, unexplained infertility, endometrial polyp. Optimum mass spectrometry techniques, determined using steroid standards, were LC/MS for androgens and LC/MS/MS for oestrogens, following dansyl-chloride derivatisation. GC/MS was less sensitive. The route of testosterone metabolism varied in the cell lines and clinical biopsies, the major metabolite was correlated with high expression of 5AR1 or 17p-HSD2, 4 and 8. If DHT (dihydrotestosterone) was produced high basal expression of 5AR1 was observed and if androstenedione was produced high basal expression of HSD2, 4 or 8 was observed. A mixture of DHT and ... (continues)