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. 2022 Nov 29;14(23):5900. doi: 10.3390/cancers14235900

Figure 2.

Figure 2

Acetyl-CoA diminishes the inhibitory effect of abiraterone and enzalutamide on AR signaling. (A) LNCaP cells were transfected with pGL3-AR-luc and phRL-TK plasmids and cultured under androgen-depleted conditions for 24 h followed by treatment with R1881 (1 nM) and the indicated concentrations of acetyl-CoA for 16 h. Samples were assayed for firefly and renilla luciferase activities using the Dual-Glo Luciferase assay. Values were normalized to Renilla activities. (B) LNCaP cells were cultured under androgen-depleted conditions for 24 h followed by treatment with R1881 (1 nM) and acetyl-CoA (0.5 mM) for 6 h. Gene expression was assayed by qRT-PCR. The 18S gene was used for normalization. (C) LNCaP cells were transfected with pGL3-AR-luc and phRL-TK plasmids and cultured under androgen-depleted conditions for 24 h followed by treatment with enzalutamide (Enz) (1 μM), abiraterone acetate (5 μM) (Abi), R1881 (1 nM), sodium acetate (10 mM), and acetyl-CoA (0.1 mM) for 16 h. (D) LNCaP and C4-2B PC cells were cultured under androgen-depleted conditions for 24 h followed by treatment with acetyl-CoA (0.1 mM) for 48 h. Expression of testosterone in cell lysates and cell culture supernatants was examined using a testosterone ELISA kit (Cayman Chemical). (E) Aliquots of LNCaP cells described in Panel C were pre-treated with or without p300/CBP inhibitors A-485 (1 μM) or C646 (25 μM). Results are expressed as the mean (n = 3) ± SD. ** p < 0.001; *** p < 0.0001; ns—non-significant.