Beas 2b

The lung cancer cell lines 95D, A549, H1299 and the human bronchial epithelial cell line BEAS-2B were obtained from American Type Culture Collection (Manassas, VA, USA).
BEAS-2B cells were treated with NNK (Sigma-Aldrich, Saint Louis, MO, USA) at a concentration of 100 mg/L. After 24 h of exposure, the culture medium was discarded, and the cells were washed twice with PBS (PBS) and grown in fresh complete culture medium. When the cells were 80% confluent, the cells were passaged. When the cells were 70% confluent, the above NNK exposure steps were repeated and the cells were passaged to the sixth generation. The NNK-induced malignant transformation of the BEAS-2B cells was detected using the soft-agar clone formation rates. And these cells were designated as 2B-NNK cells.
BEAS-2B and 2B-NNK cells were grown in bronchial epithelial basal medium (BEBM; Clonetics, Basel, Switzerland) supplemented with SingleQuots (Clonetics). 95D and H1299 cells were grown in RPMI-1640 medium (HyClone, Logan, UT, USA) with 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Waltham, MA, USA). A549 cells were grown in F-12K medium (HyClone) supplemented with 10% FBS. All cells were incubated at 37°C in a humidified incubator containing 5% CO2.

Human NSCLC cell lines (NCI‐H650, NCI‐H1299, NCI‐H1437, and A549) and human normal lung bronchus epithelial cell line BEAS‐2B were purchased from American Typical Cell Collection. The NCI‐H650, NCI‐H1299, and NCI‐H1437 cells were maintained in 90% Roswell Park Memorial Institute (RPMI) 1640 Medium (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco); the A549 cells were cultured in 90% Ham's F‐12K (Kaighn's) Medium (Gibco) supplemented with 10% FBS (Gibco). And the BEAS‐2B cells were cultured in Bronchial Epithelial Cell Growth Basal Medium (Clonetics). All cells were incubated in a humidified atmosphere at the condition of 95% air, 5% CO₂ and 37°C. After culture, circ‐SMARCA5 expression in the cells was detected by RT‐qPCR where BEAS‐2B cells were served as control.