H1915

Human LUAD cell lines H1299, H1915, H1650, A549, H1975, H661, H827, and PC-9, and the normal lung epithelial cell line HBE were all obtained from American Type Culture Collection (ATCC). All culture media were supplemented with 10% fetal bovine serum (FBS, PAN, Biotech GmbH, Germany). PC-9 was cultured in Dulbecco's Modified Eagle's Medium (DMEM, Gibco^®, Grand Island, NY, USA), while the remaining cell lines were cultured in RPMI-1640 (Gibco^®) at 37 °C in a humidified atmosphere containing 5% CO2. Cells used in experiments were in good condition without mycoplasma contamination.

Human NSCLC cell lines (PC9, A549, H1915, H1975, H1650, and H1299) and human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (Manassas, VA, USA). Human bronchial epithelial cells (HBE) and human astrocytes (HA) were purchased from the Cell Bank of Academia Sinica (Shanghai, China). A549, A549-BrM3, H1915, H1915-BrM3, H1975, H1650, H1299, HUVEC and HBE cells were cultured in RPMI-1640, and PC9 and HA cells were maintained in DMEM (HyClone, Logan, UT, USA). These media were supplemented with 10% fetal bovine serum (Gemini, Bridgewater Township, NJ, USA), 100 μg/mL streptomycin, and 100 U/mL penicillin.
Matched samples of NSCLC, normal lung tissue, and brain metastases were obtained from surgical patients at the Harbin Medical University Cancer Hospital. Detailed explanation of the research program was given to patients and informed consent was signed. Both NSCLC and brain metastasis samples were confirmed histologically. See Supplementary Table 1. for details. The GEO GSE74706 dataset was used for data validation through correlation analysis. We used the UALCAN online tool (http://ualcan.path.uab.edu/ind ex.html) to analyze the expression of HOXB9 in NSCLC in TCGA database. The present study was approved by the Ethics Committee of Harbin Medical University Cancer Hospital.

The primary glioblastoma cell line U-87 MG, the hormone-refractory prostate cancer cell line DU 145, and the large cell lung cancer cell line NCI-H1915 (hereinafter “H1915”) were purchased from the American Type Culture Collection (ATCC, Manassas, VA). DU 145 and H1915 cell lines were originally isolated from a brain metastasis. All cell lines were maintained according to the supplier’s instructions and routinely tested for mycoplasma contamination using MycoAlerTM (Lonza, Walkersville, MD, USA).