Journal: Molecular Oncology

Article Title: TAS 3681, an androgen receptor antagonist, prevents drug resistance driven by aberrant androgen receptor signaling in prostate cancer

doi: 10.1002/1878-0261.13641

Figure Lengend Snippet: TAS3681 inhibits AR function and downregulates AR. (A) Chemical structure of TAS3681. (B) Effect of TAS3681, enzalutamide, and bicalutamide on the transcriptional activity of the wild‐type AR in COS‐7 cells (upper panel) or the T878A mutant AR in LNCaP cells (lower panel). COS‐7 cells were transfected with pGL4.36 luciferase (containing androgen‐dependent murine mammary tumor virus long terminal repeat), Renilla luciferase plasmids, and AR expression vector, and incubated for 1 day. LNCaP cells were transfected with the pGLPE luciferase plasmid and incubated for 1 day. COS‐7 cells and LNCaP cells were treated with various concentrations of TAS3681, enzalutamide, or bicalutamide with 1 n m DHT for COS‐7 or with 0.1 n m DHT for LNCaP, respectively, in steroid‐depleted medium for 1 day before luciferase activity measurements. Results are expressed as mean of triplicate wells ± SD. The IC 50 values are presented in Table . (C) Inhibitory effect of TAS3681, enzalutamide, and bicalutamide on DHT‐induced proliferation of VCaP (left panel) and LNCaP cells (right panel). VCaP cells were treated with various concentrations of TAS3681, enzalutamide, and bicalutamide in the presence of DHT for 3 days. The dashed line indicates cell viability in the absence of DHT. Results are expressed as mean of sextuplicate wells ± SD. (D) AR and GAPDH protein expression in LNCaP cells treated with TAS3681, enzalutamide, and TOK‐001. LNCaP cells were treated with either the test compounds or DMSO for 1 day. Cell lysates were subjected to immunoblotting for AR and GAPDH to confirm equal loading. The numbers under each blot represent the relative expression value of the AR protein relative to that of DMSO‐treated cells. TOK‐001 was used as a positive control for the reduction in AR expression. A representative experiment (of 2) is shown. AR, androgen receptor; Enz, enzalutamide; TOK, TOK‐001.

Article Snippet: Darolutamide was obtained from MedChemExpress Co., Ltd. (Monmouth Junction, NJ, USA), TOK‐001 was obtained from Sundia MediTech Co., Ltd. (Shanghai, China).

Techniques: Activity Assay, Mutagenesis, Transfection, Luciferase, Virus, Expressing, Plasmid Preparation, Incubation, Western Blot, Positive Control