22rv1 pca

LnCaP and 22rv1 PCa cell lines obtained from ATCC (Manassas, VA, USA) were kindly donated by Dr. Guillermo Velasco. PC3 and DU145 cell lines were obtained from ATCC (Manassas, VA, USA). LnCaP, 22rv1 and DU145 were cultured in RPMI 10% FBS. PC3 were cultured in RPMI/F12 10% FBS. HEK-293 T cells cultured in 10% FBS DMEM medium were used for virus amplification. To trigger HGK signaling cells were treated with TNF-α (20 ng/mL; R&D, Minneapolis, MN, USA). The chemical inhibitor used to inhibit HGK was PF 06,260,933 (1–10 µM; Sigma, PZ0272).

LNCaP, PC3, C4-2 and 22RV-1 PCa cells were purchase from ATCC and maintained in Tissue and Cell Culture Core Facility at Baylor College of Medicine. Cells were cultured in RPMI1640 medium with 10% FBS and antibiotics (100 μg ml⁻¹ streptomycin and 100 U ml⁻¹ penicillin G) in a humidified atmosphere of 5% CO₂ and 95% air at 37 °C. LNCaP-abl cells were cultured in RPMI1640 medium with 10% charcoal-stripped FBS. Fresh medium were changed after three days of incubation. Cells were routinely checked for mycoplasma contamination by using MycoAlert Mycoplasma Detection Kit (LONZA). Short-tandem repeat analysis was performed by the service of Core Facility in MD Anderson Cancer Center for the authentication of the cell lines. For the enzalutamide experiment, 2 × 10³ LNCaP parental and enzalutamide-resistant cells were plated into 96-well-plates and treated with 10 μM enzalutamide (purchased from Selleck Chemicals) for the indicated time points.