Establishment and Characterization of Lung Cancer Cell Lines

A human lung adenocarcinoma cell line, A925L, established from a surgical specimen obtained from a Japanese male patient (T2N2M0, stage IIIA), was maintained in RPMI-1640 medium, supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (10 μg/mL), in a humidified CO₂ incubator at 37°C. The characteristics of this cell line are documented in a previous report.10 (link) The H2228 human lung adenocarcinoma cell line, with the EML4-ALK fusion protein variant 3 (E6;A20) were purchased from the ATCC (Manassas, VA, USA). The H3122 human lung adenocarcinoma cell line, with the EML4-ALK fusion protein variant 1 (E13;A20), was kindly provided by Dr. Jeffrey A. Engelman of the Massachusetts General Hospital Cancer Center (Boston, MA, USA).11 (link) PC-9 cells, an EGFR mutant human lung adenocarcinoma cell line, were obtained from Immunobiological Laboratories Co. (Fujioka, JAPAN), Ltd. All cells were passaged for <3 months before renewal from frozen, early-passage stocks. Cells were regularly screened for mycoplasma by using MycoAlert Mycoplasma Detection Kits (Lonza, Rockland, ME, USA). Crizotinib and alectinib (Fig. S1) were obtained from Selleck Chemicals (Houston, TX, USA).