SW1736 and 8505C cells, derived from human ATC, were cultured in RPMI-1640 medium (Euroclone S.p.A, Milano, Itlay) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), 2 mM L-glutamine (Euroclone S.p.A) and 50 mg/mL gentamicin (Gibco; Thermo Fisher Scientific, Inc.). Cells were cultured in a humidified incubator (5% CO2 and 95% air at 37 °C) (Eppendorf AG, Hamburg, Germany). Both cell lines were validated using short tandem repeat analysis and confirmed to be mycoplasma-free. SW1736 and 8505C cells were treated with DMSO (vehicle; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), DHT (purity ≥ 99%; Selleck Chemicals, Houston, TX, USA) or Paclitaxel (Selleck Chemicals). The DHT was prepared by dissolving 2 mg of powder in 2 mL of DMSO, resulting in a 3.6 mM stocking solution.
To obtain ATC Paclitaxel-resistant cell lines (SW1736-PTX and 8505C-PTX), the two parental cell lines were treated with increasing doses of Paclitaxel arising from 1 nM to 1.5 μM for 6 months. Resistance to pacitaxel was considered to be acquired when a 1.5 μM dose resulted in no effect in terms of cell viability. Resistant cells were cultured in RPMI-1640 medium supplemented with 10%, 2 mM L-glutamine, 50 mg/mL gentamicin and Paclitaxel 1.5 μM.
To obtain ATC Paclitaxel-resistant cell lines (SW1736-PTX and 8505C-PTX), the two parental cell lines were treated with increasing doses of Paclitaxel arising from 1 nM to 1.5 μM for 6 months. Resistance to pacitaxel was considered to be acquired when a 1.5 μM dose resulted in no effect in terms of cell viability. Resistant cells were cultured in RPMI-1640 medium supplemented with 10%, 2 mM L-glutamine, 50 mg/mL gentamicin and Paclitaxel 1.5 μM.