Hepatocyte growth factor (HGF) suppresses FSH-dependent estradiol-17β (E₂) 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 PKBα mRNA expression. Granulosa cells were incubated with androstenedione and FSH, HGF, and/or dibutyryl-cAMP (Bu₂-cAMP). Follicle-stimulating hormone and Bu₂-cAMP each stimulated estrone (E₁) and E₂ synthesis at 48 h. Hepatocyte growth factor suppressed FSH-dependent E₂, but not E₁, synthesis. Semiquantitative reverse transcription–polymerase chain reaction showed that HGF impaired FSH-supported 17β-hydroxysteroid dehydrogenase type-1 (17β-HSD) and cytochrome P450 aromatase (P450arom) mRNA levels. Hepatocyte growth factor did not reduce E₂ synthesis or 17β-HSD and P450arom mRNA expression in the presence of Bu₂-cAMP at 48 h. The FSH and HGF each down-modulated c-Met mRNA accumulation, whereas Bu₂-cAMP increased c-Met mRNA content. Between 0 and 48 h a biphasic change in PKBα mRNA content occurred with either FSH or HGF; however, PKBα mRNA accumulation was augmented by HGF. Collectively, results suggest that HGF can suppress E₂ production in GC by disrupting cAMP-dependent 17β-HSD and P450arom. Changes in c-Met and PKBα mRNA content provide a potential link between HGF signaling and the FSH-dependent mechanisms that control the steroidogenic differentiation of GC.