Figure 4
DHT inhibited cell proliferation through regulating p-Akt expression by enhancing PTEN and suppressing resultant cyclin D1 expression.
The data are means ± SD. (A) Rat granulosa cells were pre-treated with different doses of DHT for 1 hour prior to FSH treatment and after 24 hours, cell lysates were used to measure the cyclin protein levels by western blot. Expression of tubulin was used as loading control. Representative immunoblottings of protein are shown. N = 3. Relative density (RD) of each lane was determined by ImageJ, lane 1 was defined as 1. Statistical comparisons were done between lane 1 with the other lane. *P < 0.05. (B) Rat granulosa cells were pre-treated with DHT (50 μM) or insulin (500 nM) for 1 hour prior to FSH (10 ng/ml) treatment and after 24 hours, cell lysates were used to measure the indicated protein level by western blot. Expression of β-actin was used as loading control. Representative immunoblottings of protein are shown. N = 3. ImageJ determined RD and lane 1 was defined as 1. Statistical comparisons were done between lane 1 with the other lane. *P < 0.05. (C) Rat granulosa cells were pre-treated with DHT or DHEA and after 24 hours, cell lysates were used to measure the PTEN level by western blot. Expression of tubulin was used as loading control. Representative immunoblottings of protein are shown. N = 3. ImageJ determined RD and lane 1 was defined as 1. Statistical comparisons were done between lane 1 with the other lane. *P < 0.05. (D) Rat granulosa cells were pretreated with PTEN siRNA(si) or control siRNA(25 μM) for 24 hours and following with or without FSH(10ng/ml) and DHT(50 μM) treatment. After 3 days, cell growth was determined by the MTT assay, N=3. Between-group comparisons were performed as indicated *P < 0.05.
