Expression of 3b-Hydroxysteroid dehydrogenase and P450 side chain cleavage enzyme in the human uterine endometrium

The enzyme complex 3b-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3beta-HSD) is involved in the biosynthesis of all classes of active steroids. The expression of 3beta-HSD in human uterine endometrium during the menstrual cycle and decidua was examined in an effort to understand its role during ova implantation. 3beta-HSD was weakly expressed in the glandular epithelium of the proliferative phase and moderately expressed in the glandular epithelium of secretory phase of the endometrium. In the decidua of the ectopic pregnancy, 3beta-HSD was strongly expressed. The human uterine endometrial 3beta-HSD was identified as being the same type as the placental 3beta-HSD by RT-PCR and sequence analysis. In addition to the expression of 3beta-HSD, P450scc was expressed in the decidua of the ectopic pregnancy. These results suggest that pregnenolone might be synthesized from cholesterol by P450scc de novo and then, it is converted to progesterone by 3beta-HSD in the uterine endometrium. The data implies that the endometrial 3beta-HSD can use not only the out-coming pregnenolone from the adrenal gland but also the self- made pregnenolone to produce progesterone. The de novo synthesis of progesterone in the endometrium might be a crucial factor for implantation and maintenance of pregnancy.

Diagnostic Significance of 3beta-Hydroxysteroid Dehydrogenase Polyclonal in Human Steroidogenic Tumors / 대한해부학회지

The enzymatic complex 3beta-hydroxysteroid dehydrogenase Delta5-Delta4 isomerase(3beta-HSD) catalyzes the obligatory oxidation and isomerization of Delta4-hydroxypregnene and Delta5-hydroxyandrostene steroid precursors into Delta4-ketosteroid necessary for the formation of all classes of steroid hormones such as glucocorticoid, mineralocorticoid, progesterone, androgen and estrogen. The 3beta-HSD enzymatic activity is present at the syncytiotrophoblast, the sebaceous gland of the skin, the Leydig cells of the testis, the thecal cells and granulosa lutein cells ofthe ovary in human. So we thought that 3beta-HSD antibody can be used to diagnose several steroidogenic tumors in human, but there has been few trial for it. In this study, we made polyclonal antibody against human placental 3beta-HSD. And then immunohistochemical studies with the antibody were performed to diagnose several kinds of steroidogenic tumors in human. The polyclonal antibody detected specifically the placental protein of 43 KDa by Western blotting. And the polyclonal antibody reacted on the syncytiotrophoblast, the sebaceous gland of the skin, the Leydig cells of the testis, adrenal cortex, the thecal cells and granulosa lutein cells of the ovary with immunohistochemical staining. In immunohistochemical staining, the polyclonal antibody against 3beta-HSD reacted on the primary and metastatic choriocarcinoma, placental site trophoblastic tumor, sebaceous carcinoma, adrenal cortical adenoma and carcinoma. But it did not react on the squamous cell carcinoma, the basal cell carcinoma and malignant melanoma of the skin, the adenocacinoma and the leiomyosarcoma of the uterus, the metastatic and primary renal cell carcinoma. From these results, the polyclonal antibody against human placental 3beta-HSD demonstrated to have specific reaction on steroidogenic tumors and also it can be used in differential diagnosis of primary and metastatic related to steroid hormone.

Molecular cloning and expression of 3beta-hydroxy steroid dehydrogenase in E-Coli / 대한해부학회지

This study was carried out to obtain the 3beta-hydroxy-5-ene steroid dehydrogenase (3beta-HSD) which is contained in human trophoblasts. To produce 3beta-HSD cDNA, reverse transcription-polymerase chain reaction (RT-PCR) with the total RNA of human placenta was performed and RT-PCR products was detected at 1.2 Kb. After cloning 3beta-HSD cDNA in pGEX 4T-2 as expression vector, the recombinant DNA was transformed to E-coli and glutathione-s-transferase (GST) fusion protein was inducted with isoprophyl thiogalactoside (IPTG). Inducted GST fusion protein was overexpressed in cytoplasm of E-coli. The obtained GST fusion protein was characterized with antibody against 3beta-HSD by Western blot analysis and purified by affinity chromatography using glutathione sepharose 4B column. And then the GST fusion protein was cleaved by thrombin and The cleaved protein, 3beta-HSD, was detected at 43 KDa by SDS-PAGE.

The Localization of 3beta-hydroxysteroid Dehydrogenase/Delta5-Delta4 isomerase in the Cardiovascular System of Rats / 대한해부학회지

The enzyme complex 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is involed in the biosynthesis of all classes of active steroids. It has been known that the enzymatic activity of 3beta-HSD is present not only in the classical steroido-genic tissues, but also in many peripheral tissues. This study was performed to investigated of 3beta-HSD immunore-activity in the rat cardiovascular tissues such as the ventricle, atrium, aortic arch, and abdominal aorta. Immunoblot analyses and immunohistochemical studies were performed using polyclonal antibodies raised against purified human placental 3beta-HSD. We identified 43 and 37KDa bands in the ventricle and atrum, whereas only 37KDa band was observed in the aortic arch and abdominal aorta. Immunostaining for 3beta-HSD was detected in the ventricular and atrial cardiocytes. The intensity of staining was much higher in the atrial cardiocytes than in the ventricular cardiocytes. Immunostaining was also found in the smooth muscles of aortic arch and abdominal aorta.