Figure 5.

Effect of leptin on aromatase mRNA expression and diminished expression of Sam68 and aromatase in GCs from PCOS women. GCs samples were obtained from 25 healthy donors and 25 PCOS women. GCs were cultured in a medium containing 0% FCS for 24 h. (A) GCs from healthy donors were treated with or without (control) 10 nM leptin for 24 h. (B) GCs from PCOS women were treated with or without (control) 10 nM leptin for 24 h. Aromatase gene expression was analyzed by qRT-PCR, normalized against RPS-7 and compared to the control group. (C) Representative Western blot analysis of Sam68 protein level in GCs from healthy donors and PCOS women. GCs lysates were denatured and resolved by SDS-PAGE with anti-Sam68 antibodies. Loading controls were performed in the same membranes with anti-GAPDH. We show the corresponding densitometric analysis of three independent experiments as means ± s.d. Statistical analyses were performed by ANOVA. (D) Relative mRNA level of Sam68 in GCs from healthy donors and PCOS women. Sam68 mRNA was quantified with qRT-PCR. Cyclophylin was used as internal standard. Paired t-test was performed to examine the difference in mRNA level of Sam68 and statistical significance was considered when P value was < 0.05. (E) Relative mRNA level of aromatase in GCs from healthy donors and PCOS women, quantified by qRT-PCR. RNA was extracted as described in Materials and Methods. RPS-7 was used as internal standard. Paired t-test was performed to examine the difference in mRNA level of aromatase, and statistical significance was considered when P value was < 0.05. Asterisks indicate significant differences from the control according to Mann–Whitney U test.