Cell Culture of LUAD and Control Cell Lines

LUAD cell lines (A549, H1975, and CALU-3) were cultured in RPMI 1640 medium (Gibco, Waltham, MA, USA), and human bronchial epithelioid cell line (HBE) and HEK293T cell lines were cultured in Dulbecco’s modified eagle medium (Gibco). The cell culture medium was supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (New Cell & Molecular Biotech, Suzhou, China). All cells were obtained from the cell bank of the Cancer Research Institute of Central South University (Changsha, China) [21 (link),22 (link)] and incubated at 37 °C with 5% CO₂. The cisplatin-resistant A549 cell line (A549/CDDP) was derived from A549 cells using stepwise increased cisplatin (Qilu Pharmaceutical, Shandong, China) concentration incubation. All cells were passaged to the third generation to start the experiments.