title: Modulation of estrogen production and 17beta-hydroxysteroid dehydrogenase-type 1, cytochrome P450 aromatase, c-met, and protein kinase Balpha messenger ribonucleic acid content in rat ovarian granulosa cells by hepatocyte growth factor and follicle-stimulating hormone.
abstract: Hepatocyte growth factor (HGF) suppresses FSH-dependent estradiol-17beta (E(2)) production in ovarian granulosa cells (GC). The mechanisms of action for HGF in GC are unknown; however, activation of the HGF receptor, c-Met, can induce c-Akt/protein kinase B (PKB)-mediated signal transduction in nonovarian cells. Using immature rat GC, the present study investigated the effects of HGF within the estrogen biosynthetic pathway, concomitant with changes in c-Met and PKBalpha mRNA expression. Granulosa cells were incubated with androstenedione and FSH, HGF, and/or dibutyryl-cAMP (Bu(2)-cAMP). Follicle-stimulating hormone and Bu(2)-cAMP each stimulated estrone (E(1)) and E(2) synthesis at 48 h. Hepatocyte growth factor suppressed FSH-dependent E(2), but not E(1), synthesis. Semiquantitative reverse transcription-polymerase chain reaction showed that HGF impaired FSH-supported 17beta-hydroxysteroid dehydrogenase type-1 (17beta-HSD) and cytochrome P450 aromatase (P450arom) mRNA levels. Hepatocyte growth factor did not reduce E(2) synthesis or 17beta-HSD and P450arom mRNA expression in the presence of Bu(2)-cAMP at 48 h. The FSH and HGF each down-modulated c-Met mRNA accumulation, whereas Bu(2)-cAMP increased c-Met mRNA content. Between 0 and 48 h a biphasic change in PKBalpha mRNA content occurred with either FSH or HGF; however, PKBalpha mRNA accumulation was augmented by HGF. Collectively, results suggest that HGF can suppress E(2) production in GC by disrupting cAMP-dependent 17beta-HSD and P450arom. Changes in c-Met and PKBalpha mRNA content provide a potential link between HGF signaling and the FSH-dependent mechanisms that control the steroidogenic differentiation of GC.
authors.affiliation: Southern Illinois University (SIU) School of Dental Medicine, Department of Applied Dental Medicine, Alton, Illinois 62002-4700, USA. rzachow@siue.edu
authors.forename: R J
authors.initials: RJ
authors.lastname: Zachow
authors.forename: B E
authors.initials: BE
authors.lastname: Ramski
authors.forename: H
authors.initials: H
authors.lastname: Lee
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000235:genetics; D000818:Animals; D001141:Aromatase / Q000235:genetics; D003994:Bucladesine / Q000494:pharmacology; D004958:Estradiol / Q000096:biosynthesis; D004967:Estrogens / Q000096:biosynthesis*; D004970:Estrone / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000737:chemistry / Q000378:metabolism*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000235:genetics; D051057:Proto-Oncogene Proteins c-akt; D019859:Proto-Oncogene Proteins c-met / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D004967:Estrogens; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D004970:Estrone; D004958:Estradiol; D003994:Bucladesine; D017228:Hepatocyte Growth Factor; D005640:Follicle Stimulating Hormone; D015067:17-Hydroxysteroid Dehydrogenases; D001141:Aromatase; D019859:Proto-Oncogene Proteins c-met; C494920:Akt1 protein, rat; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt