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Fig. 4 | Cell & Bioscience

Fig. 4

From: Anti-metastatic effect of GV1001 on prostate cancer cells; roles of GnRHR-mediated Gαs-cAMP pathway and AR-YAP1 axis

Fig. 4

Involvement of Gαs-cAMP signaling pathway in GV1001-stimulated AR activation. A Effects of Gα signaling inhibitors on GV1001-stimulated AR phosphorylation in LNCaP cells. LNCaP cells were exposed to various Gαs inhibitors, a cell-permeable cytosolic calcium chelator BAPTA/AM (10 μM), a specific adenylyl cyclase inhibitor KH7 (5 μM) and an ADP-ribosyltransferase of Gαi, pertussis toxin (100 ng/ml) with 10 μM GV1001. All of the results were confirmed by multiple experiments. B Inhibitory effect of KH7 on GV1001-induced ARE-reporter activity. LNCaP cells were transiently transfected with ARE-luciferase plasmid (1 μg/ml) and the cells were exposed to KH7 (5 μM) and GV1001 (10 μM) for 24 h. Luciferase activity was determined to assess ARE responsiveness. Data represent means ± SD (n = 5, **P < 0.01 vs vehicle-treated control, ###P < 0.005 vs GV1001-treated group). C AR activation by forskolin. LNCaP cells were treated with forskolin, a cAMP activator (0.1–10 μM) for 24 h, and protein expression of AR and phosphorylated AR was assessed by immunoblotting. D Inhibitory effect of KH7 on forskolin-stimulated AR expression and phosphorylation. LNCaP cells were exposed to 5 μM KH7 and 10 μM forskolin for 24 h. E Inhibitory effect of KH7 on forskolin-induced ARE-reporter activity in LNCaP cells. LNCaP cells were transiently transfected with ARE-luciferase plasmid (1 μg/ml) and the cells were exposed to KH7 (5 μM) and forskolin (10 μM) for 24 h. Luciferase activity was determined to assess ARE responsiveness. Data represent means ± SD (n = 6, **P < 0.01 vs vehicle-treated control, ##P < 0.01 vs forskolin-treated group). F, G Inhibitory effect of LA on DHT-induced AR activation. F Inhibitory effect of LA on DHT-induced AR phosphorylation. LNCaP cells were pretreated with LA (100 nM) for 1 h and treated with DHT (100 nM) for 24 h. G Inhibitory effect of LA on DHT-induced ARE-reporter activity. LNCaP cells were transiently transfected with ARE-luciferase plasmid (1 μg/ml) and the cells were exposed to DHT (100 nM) and LA (100 nM) for 24 h. Luciferase activity was determined to assess ARE responsiveness. Data represent means ± SD (n = 6, **P < 0.01 vs vehicle-treated control, ###P < 0.005 vs DHT-treated group). H Hypothetical diagram for the differential effects of GV1001 and LA on AR

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