Human wtEGFR NSCLC cell lines H358 and A549 were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). Cisplatin-resistant cell lines, H358R and A549R, were induced by constant exposure to cisplatin (2 μmol/L) to imitate acquired resistance. Both cell lines were cultured in 10% FBS-containing medium (RPMI1640, Gibco, Thermo Fisher Scientific) and maintained in a 5% CO2 incubator at 37 °C. Cisplatin (A8321) was purchased from APExBIO Technology LLC (Houston, Texas, USA); Gefitinib (ZD1839) was obtained from Med Chem Express (Monmouth, Junction, USA). (3, 4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-benzimidazolyl carbocyanine iodide JC-1 were provided by Sigma-Aldrich (St. Louis, MO, USA). Antibodies used were following: total EGFR (#4267), phospho-EGFR (#3777), Phospho-AKT Antibody Sampler Kit (#9916), Phospho-Erk1/2 Antibody Sampler Kit (#9911), Apoptosis Antibody Sampler Kit (#9915) and other antibody sampler kits were obtained from Cell Signaling Technology (Danvers, MA, USA).
Gefitinib zd1839
Manufactured by MedChemExpress
Sourced in United States
Gefitinib (ZD1839) is an FDA-approved small-molecule tyrosine kinase inhibitor. It functions by selectively inhibiting the epidermal growth factor receptor (EGFR) tyrosine kinase.
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Lab products found in correlation
Phospho egfr, by Cell Signaling Technology (1 mentions)
Phospho akt antibody sampler kit, by Cell Signaling Technology (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Total egfr, by Cell Signaling Technology (1 mentions)
Apoptosis antibody sampler kit, by Cell Signaling Technology (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hcc827, by American Type Culture Collection (1 mentions)
Nci h1975, by American Type Culture Collection (1 mentions)
2 protocols using gefitinib zd1839
1
Cisplatin-Resistant NSCLC Cell Lines
2
Cell Line Development and Characterization
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Human normal lung epithelial cell line HBE (ATCC CRL-2078), human NSCLC cell line NCI-H1975 (EGFRL858R+T790M, gefitinib-resistant, ATCC CRL-5908), and HCC827 (ATCC CRL-2868) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA), while human NSCLC cell line PC9 (EGFRexon19 del; gefitinib-sensitive) was kindly provided by Shenzhen People’s Hospital (Shenzhen, China). Gefitinib-resistant HCC827 (HCC827GR) cells were established at increasing concentrations of gefitinib for approximately five months. These cell lines were cultured at 37 °C in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37 °C in a humidified atmosphere of 5% CO2. Fasudil (HA-1077) and gefitinib (ZD1839) were purchased from MedChem Express (Monmouth Junction, NJ, USA). FBS, antibiotics, and RPMI-1640 medium were purchased from Gibco (Carlsbad, CA, USA).
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