Beas 2b

The human bronchial epithelial cell lines, NL-20 (BCRC) and BEAS-2B (ATCC), were cultured in LHC-9 medium (Invitrogen, NY, USA) supplemented with 5% fetal bovine serum, 100 U/mL penicillin, and 100 pg/mL streptomycin (Invitrogen) in a humidified incubator with 5% CO₂ at 37 °C.

Human lung epithelial cell line BEAS-2B, NSCLC cell lines (H1299/A549/PC9/H292), and HEK293T cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). DMEM medium (C11995500BT; Gibco) was used to culture HEK293T cells. RPMI 1640 medium (C11875500BT; Invitrogen) was used to culture BEAS-2B, H1299, A549, PC9, and H292 cells. The cells were fed in a medium containing 10% fetal bovine serum (10091148; Gibco) and placed in an incubator containing 5% CO₂ at 37 °C. The cells were cultured in six-well plates until the density approximately reached 70–80%. Then, the plasmids or siRNAs were transfected into cells according to the instructions of the transfection reagent kit (2102-100/2103-100; Pufei).

Two pulmonary epithelial cell types were used in the present study. All cells were maintained at 37 °C and 5% CO₂ with standard aseptic procedures. Immortalized human bronchial epithelial cells (BEAS-2B, ATCC, Manassas, VA) of less than 10 passages upon arriving in our laboratory were used to examine cytotoxicity, nuclear uptake, cell cycle arrest, mitotic aberrations, centrosome integrity, and spindle pole integrity. BEAS-2B were cultured in Dulbecco’s Modified Eagle Medium (DMEM) media supplemented with 10% (v/v) serum (Invitrogen, Grand Island, NY) and 1% (v/v) antibiotic-antimycotic (Corning, Corning, NY). Primary small airway respiratory epithelial cells (SAEC; Lonza, Walkersville, MD) from a non-smoking human donor were used to examine cytotoxicity, nuclear uptake, cell cycle arrest, aneuploidy, and clonal growth. The normal karyotype of the primary cells was essential for the examination of aneuploidy. The SAEC were cultured following manufacturer’s directions and using Cabrex media (Lonza, Walkersville, MD). Epithelial phenotype was identified in both cell types through EM analysis of stained cytokeratin 8 and 18 (data not shown) [37 (link)].

Human melanoma cell line SK-MEL-2, human breast cancer cell line MDA-MB-231, and human lung cancer cell line A549 were maintained in DMEM (Invitrogen) while acute myeloid leukemia cell line THP-1, Non-small cell lung cancer cell line H1299, and normal lung epithelial cell line BEAS-2B were cultured in RPMI (Invitrogen). All cancer cell lines were obtained from Bioresource Collection and Research Center (BCRC), Taiwan. BEAS-2B was a gift from Dr. I-Ching Wang of National Tsing Hua University (Taiwan). All culture media were supplemented with 10% fetal bovine serum (FBS; Biological Industries, Israel or Hyclone, USA) and 1% of penicillin-streptomycin (GIBCO), cultured at 37°c with 5% CO₂.