Du145 htb 81

Prostate cancer (DU145 (HTB-81)), head and neck squamous cell carcinoma (Fadu (HTB-43)), pancreatic cancer (BxPC3 (CRL-1687)), melanoma (A375M (CRL-3222), MDA-MB-435 (HTB-131), Sk-mel 24 (HTB-71), Sk-mel 28 (HTB-72), WM115 (CRL-1675)), cells were purchased from American Type Culture Collection (Rockville, MD, USA). Ovarian cancer (OVCAR-8) were from the National Cancer Institute/National Institutes of Health (NCI/NIH; Frederick, MD, USA). Gastric cancer cells (SNU638) were purchased from the Korean Cell Line Bank (KCLB). All cell lines were cultured less than 3 months after resuscitation. The cells were cultured according to manufacturers’ instructions, using EMEM (Eagle’s Minimum Essential Medium) from Thermo Fisher Scientific, Inc., (Waltham, MA, USA) for DU145, Fadu, Sk-mel 28, Sk-mel 24 and WM115 cells, DMEM (Dulbecco’s Modified Eagle’s Medium) from Thermo Fisher Scientific, Inc. for A375M, and RPMI-1640 medium (Thermo Fisher Scientific, Inc.) for MDA-MB-435, OVCAR-8 and SNU638 cells, supplemented with 10% heat-inactivated fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA), l-glutamine, 100 units/mL penicillin and 100 μg/mL streptomycin (Sigma-Aldrich Corporation, St. Louis, MO, USA), and incubated at 37 °C in humidified air with 5% CO₂.

Human PCa DU145 (HTB-81) and PC3 (CRL-1435) cell lines was obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and was received on 2016/2018. This cell line has been tested and authenticated by DNA fingerprinting by the ATCC. After reception, cells were amplified in order to make a large reserve of cryopreserved cells. Every 3 months, a new cryopreserved bulb was thawed and used for this study. DU145 and PC3 cell lines were grown in RPMI medium (BE12-702F, Lonza, Levallois-Perret, France), supplemented with 5% FBS (CVFSVF0001, Eurobio, Les Ulis, France) and 1% (v/v) penicillin–streptomycin in a humidified incubator at 37 °C with 5% CO₂. All experiments were performed with mycoplasma-free cells. DU145 and PC3 PCa cell lines have been chosen for their high migration capacity.
LA (L1876), EPA (17266), palmitic acid (PA) (P5177), and TGFβ1 (H8541) were from Sigma-Aldrich. GSK7975A (GLXC03243) and Synta66 (GLXC03244) were from GLXX Lab IMC. 1-Ohexadecyl-2-O-methyl-sn-glycero-3-lactose (Ohmline) was synthetized as previously described [33 (link)].

LNCaP (CRL-1740), HEK 293T (CRL-11268), PC-3 (CRL-1435), PPC-1 (HTB-190) and DU145 (HTB-81) cells were purchased from the American Type Culture Collection (ATCC; Manassa, VA, USA). These ATCC cell lines were authenticated by STR analysis. C4-2 and CWR22Rv1 cells were obtained from Dr. Leland W. Chung’s laboratory (Cedars-Sinai Medical Center, Los Angeles, CA, USA), which were characterized by in vitro characterization techniques [15 (link),16 (link)]. A recombinant adenovirus E1 expressing HEK-293 (AD-293; 240085) cells were purchased from Agilent Technologies, Inc. (Santa Clara, CA, USA). LNCaP, C4-2, CWR22Rv1, and PC-3 cells were maintained in RMPI-1640 media (HyClone, Logan, UT, USA) with 5% fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA). PPC-1, AD-293, and HEK 293T cells were cultured in DMEM (HyClone, USA) with 10% FBS. Media containing 5% charcoal-stripped serum (cFBS) was used for the starvation and steroid studies. All cell lines were maintained in the presence of 100 units/mL penicillin and 100 mg/mL streptomycin (Gibco, USA) in a 5% CO₂ incubator at 37 °C.

DU145 (HTB-81) and PC3 (CRL1435) as androgen-independent prostate cancer cell lines were purchased from the American Type Culture Collection (ATCC, USA). Both cell lines were grown in DMEM containing 10% fetal bovine serum, penicillin (100 U/mL), and streptomycin (100 ng/mL) at 37°C with 5% CO₂. All the cell culture-related materials were purchased from Gibco Company, USA, unless mentioned otherwise. For hypoxia experiments, cells were grown in hypoxia incubator (Thermo Scientific, USA) in 1% oxygen for indicated time point. 20% oxygen was used as the normoxic control.