Sb202190

DA (Fig. 1) was obtained from the Sichuan Academy of Chinese Medical Sciences. The purity was 99.97 % as determined by high-performance liquid chromatography. DA was dissolved in dimethylsulphoxide (DMSO) at a concentration of 200 mmol/L. The final concentration of DMSO in the medium was not more than 0.1 % (v/v). The inhibitors PD98059 (an inhibitor of ERK), SB203580 and SB202190 (two different inhibitors of p38), SP600125 (an inhibitor of JNK) were purchased from Beyotime (Jiangsu, China). SB203580 is a pyridinyl imidazole derivative that specifically suppresses phosphorylation of p38. SB202190 is a highly potent and cell-permeable inhibitor of p38 that selectively inhibits the p38 a and b isoforms. Each inhibitor was reconstituted in DMSO and added to the culture medium so that the final concentration of DMSO was less than 0.1 % of the total volume.Fig. 1

Molecular structure of Androg (a) and DA (b)

CRPC with docetaxel chemotherapy-resistant patient-derived organoids was established pursuant to the method as described previously [51 (link),52 (link)]. The organoids were cultured with the advanced DMEM/F12 supplemented with 125 ng/ml rhR-spondin-1 (STEMCELL Technologies, 78213.1), 100 ng/ml rhNoggin (R&D Systems, 6057-NG-025), 1 ng/ml rhFGF (R&D Systems, 233-FB-025), rhFGF-10 (R&D Systems, 345-FG-025), 50 ng/ml (Meilunbio, China, MB8218-1), 1×N21 (R&D Systems, AR008), 10 μM Y-27632 (Abcam, Ab120129), 0.5 μM A83-01 (Beyotime Biotechnology, SF7917), 1:100 primocin (Invivogen, ant-pm-1), 10 μM SB202190 (Beyotime Biotechnology, SC0380), 10 mM Nicotinamide (Beyotime Biotechnology, S1761), and 1.25 mM N-acetylcysteine (Selleck, S1623), and the medium was changed every 2 to 3 days. Lentiviral transduction of AZGP1P2 shRNA and shRNA control was conducted according to the method as previously described [53 (link)]. The proliferation of organoids treated with 20 nM docetaxel was assessed by CCK-8 assay on the fifth day, and organoids were photographed using light microscopy. Three independent experiments were performed.

CRPC with docetaxel chemotherapy-resistant patient-derived organoids was established pursuant to the method as described previously [51 (link),52 (link)]. The organoids were cultured with the advanced DMEM/F12 supplemented with 125 ng/ml rhR-spondin-1 (STEMCELL Technologies, 78213.1), 100 ng/ml rhNoggin (R&D Systems, 6057-NG-025), 1 ng/ml rhFGF (R&D Systems, 233-FB-025), rhFGF-10 (R&D Systems, 345-FG-025), 50 ng/ml (Meilunbio, China, MB8218-1), 1×N21 (R&D Systems, AR008), 10 μM Y-27632 (Abcam, Ab120129), 0.5 μM A83-01 (Beyotime Biotechnology, SF7917), 1:100 primocin (Invivogen, ant-pm-1), 10 μM SB202190 (Beyotime Biotechnology, SC0380), 10 mM Nicotinamide (Beyotime Biotechnology, S1761), and 1.25 mM N-acetylcysteine (Selleck, S1623), and the medium was changed every 2 to 3 days. Lentiviral transduction of AZGP1P2 shRNA and shRNA control was conducted according to the method as previously described [53 (link)]. The proliferation of organoids treated with 20 nM docetaxel was assessed by CCK-8 assay on the fifth day, and organoids were photographed using light microscopy. Three independent experiments were performed.