FSH Stimulation promotes progesterone synthesis and output from human granulosa cells without luteinization

• Published in: Human reproduction (Oxford) 2017-03, Vol.32 (3), p.643-652
• Main Authors: Oktem, Ozgur, Akin, Nazli, Bildik, Gamze, Yakin, Kayhan, Alper, Ebru, Balaban, Basak, Urman, Bulent
• Format: Article
• Language: English
• Subjects: 3-Hydroxysteroid Dehydrogenases - genetics; 3-Hydroxysteroid Dehydrogenases - metabolism; Cell Line; Female; Follicle Stimulating Hormone - therapeutic use; Granulosa Cells - drug effects; Humans; Luteinization - drug effects; Ovulation Induction - methods; Pregnenolone - metabolism; Progesterone - biosynthesis; Progesterone - blood; RNA, Messenger - metabolism; Up-Regulation - drug effects
• ISSN: 0268-1161; 1460-2350
• DOI: 10.1093/humrep/dex010

Abstract STUDY QUESTION Can granulosa cells produce progesterone (P) in response to FSH stimulation? SUMMARY ANSWER FSH actively promotes P synthesis and output from granulosa cells without luteinization by up-regulating the expression and increasing enzymatic activity of 3β-hydroxysteriod dehydrogenoase (3β-HSD), which converts pregnenolone to P. WHAT IS KNOWN ALREADY Serum P level may rise prematurely prior to ovulation trigger in stimulated IVF cycles and adversely affect implantation and clinical pregnancy rates by impairing endometrial receptivity. STUDY DESIGN, SIZE, DURATION A translational research study. PARTICIPANTS/MATERIALS, SETTING, METHODS Human ovarian cortical samples (n = 15) and non-luteinizing FSH-responsive human mitotic granulosa cell line (HGrC1) were stimulated with rec-FSH at 12.5, 25 and 50 mIU/ml concentrations for 24 and 48 h. FSH receptor expression was knocked-down and up-regulated in the granulosa cells using short hairpin RNA (shRNA) technology and activin-A administration, respectively. The expressions of the steroidogenic enzymes were analyzed at mRNA level by real-time quantitative RT-PCR, and protein level by western blot and immunoprecipitation assay. The enzymatic activity of 3β-HSD was measured using a spectrophotometric method. In vitro estradiol (E2) and P productions of the cells before and after FSH stimulation were measured by electro-chemiluminescence immunoassay method. MAIN RESULTS AND THE ROLE OF CHANCE Stimulation of the HGrC1 cells with FSH resulted in a dose-dependent increase in the mRNA and protein level of 3β-HSD. Overall, when all time points and FSH doses were analyzed collectively, FSH significantly up-regulated the mRNA expression of its own receptor (3.73 ± 0.06-fold, P