Nci h1581

CHO-S cells (Thermo Fisher Scientific) were cultured in serum-free Power-CHO medium (Lonza) supplemented with 8 mM L-glutamine (Thermo Fisher Scientific) and 1% penicillin and streptomycin mix (Biowest). The cells were subcultered every 2–3 days at a seeding density of 0.2–0.3 × 10⁶ cells·mL⁻¹. The cells were grown at 37°C with 8% CO₂ in a shaking incubator (140 rpm).
NCI-H520 (lung squamous cell carcinoma, FGFR1-positive), NCI-H1581 (lung large cell carcinoma, FGFR1-positive), and HCC95 (lung squamous cell carcinoma, FGFR1-negative) were obtained from the American Type Culture Collection (ATCC). NCI-H520 and NCI-H1581 were cultured in RPMI 1640 (Gibco) with 10% fetal bovine serum (Biowest) and 1% penicillin and streptomycin mix (Biowest); HCC95 cells were cultured in RPMI 1640 (Gibco) with 10% fetal bovine serum (Biowest), sodium bicarbonate (Gibco), and 1% penicillin and streptomycin mix (Biowest). The cancer cell lines were cultured at 37°C with 5% CO₂.

Three human cancer cell lines were used to evaluate the antiproliferative activity of the obtained compounds: melanoma cell line Hs294T, pancreatic tumor cell line MIA PaCa-2, and lung tumor cell line NCI-H1581. The normal mouse fibroblasts cell line (BALB/3T3 clone A31) was selected as reference. All these lines were purchased from the ATCC (American Type Culture Collection, Rockville, MD, USA). NCI-H1581 cells were grown in RPMI 1640 with GlutaMax medium (Gibco, Thermo Fisher Scientific, Leicestershire, UK) with 5% fetal bovine serum (FBS; HyClone, GE Healthcare, Logan, UT, USA). Hs294T and BALB/3T3 cells were grown in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Leicestershire, UK) supplemented with 2 mM L-glutamine and 10% FBS (HyClone, GE Healthcare, Logan, UT, USA). MIA PaCa-2 cells were grown in DMEM (Gibco, Thermo Fisher Scientific, Leicestershire, UK) supplemented with 2 mM L-glutamine, 2.5% Horse Serum (Gibco, Thermo Fisher Scientific, Leicestershire, UK), and 10% FBS (HyClone, GE Healthcare, Logan, UT, USA). All culture media were supplemented with antibiotics: 100 U/mL penicillin (Polfa Tarchomin SA, Warsaw, Poland) and 100 µg/mL streptomycin (Sigma-Aldrich Chemie GmbH, Steinheim, Germany); cells were grown at 37 °C in a humid atmosphere with 5% CO₂.