Heepic

Esophageal cancer cell lines (TE1, TE3, TE5, TE6, TE8, TE10, KYSE510, and T.Tn), were purchased from the Japanese Collection of Research Bioresources Cell Bank. Esophageal epithelial cells (HEEpiC) were purchased and cultured by the supplier's protocol (ScienCell, San Diego, CA, USA). TE1, TE3, TE5, TE6, TE8, TE10, and KYSE510 were routinely propagated in RPMI 1640 (Wako, Tokyo, Japan) supplemented with 10% FBS, penicillin and streptomycin. T.Tn was propagated in DMEM/Ham's F‐12 (Wako) supplemented with 10% FBS, penicillin and streptomycin. All cell lines were maintained at 37°C, 5% CO₂ and 95% humidified air. We used 3.5‐cm NanoCulture Plate (SCIVAX, Kawasaki, Japan) for 3D culture.

Six ESCC cell lines (KYSE30, KYSE140, KYSE150, KYSE180, KYSE410, and KYSE510) were purchased from the German Culture Collection (DSMZ, Braunschweig, Germany). Human normal esophageal epithelium cells (HEEpiC) were obtained from ScienCell (Carlsbad, CA). These cells were grown in RPMI 1640 (Biowhittaker, Walkersville, MD) supplemented with 10% fetal bovine serum (HyClone, Logan, UT) at 37°C under 5% CO₂ and saturated moisture.
For investigating the role of HOTAIR, KYSE30 and KYSE510 cells were transfected with either siRNAs targeting HOTAIR (si-HOTAIR) or scrambled negative controls (si-NC; GenePharma, Shanghai, China). For investigating the relationship between HOTAIR and miR-1, HOTAIR cDNA and miR-1 mimics (GenePharma) were cloned into the mammalian expression vector pcDNA3.1 (Invitrogen, Carlsbad, CA) individually or in combination, and then the vectors were transfected into cells. For the function study of CCND1, KYSE30 and KYSE510 cells were transfected with either si-CCND1 or si-NC. All transfection reactions were performed by using Lipofectamine 2000 (Invitrogen) following the manufacturer's instructions.

Human lung cancer cell lines (H1299, 95-D, SPCA-1, A549, SK-MES-1, PC-9, H2170 and Hcc-827), human esophageal cancer cell lines (TE-11 and EC-109), human normal esophageal cell lines (HET-1A and HEEpiC), human acute monocytic leukemia cell line (THP-1), and human breast cancer cell line (MCF-7) were purchased from the Cell Bank of the China Academy of Science (Shanghai, China). THP-1, EC-109 and all the human lung cancer cell lines were grown in RPMI-1640 medium (Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gemini, Woodland, CA, USA), 100 IU/ml penicillin and 100 µg/ml streptomycin. TE-11 and MCF-7 cells were grown in Dulbeccos modified Eagles medium (DMEM) culture medium (Life Technologies) supplemented with 10% FBS, 100 IU/ml penicillin and 100 µg/ml streptomycin. HET-1A and HEEpiC were grown in EpiCM-2 complete medium (ScienCell, Carlsbad, CA, USA) with 100 IU/ml penicillin and 100 µg/ml streptomycin. All of the cells were cultured at 37°C in a humidified incubator with 5% CO₂.

The human oesophageal epithelial cell ‘HEEpiC' was obtained from ScienCell (San Diego, CA, USA) and cultured in Epithelial Cell Medium-2. Human oesophageal epithelial cells ‘Het-1A' were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in complete growth medium (Bronchial Epithelial Cell Medium (BEGM BulleKit)). Nine human oesophageal squamous cancer cell lines (TE-1, TE-5, TE-6, TE-8, TE-9, TE-10, TE-11, TE-14 and TE-15) were obtained from the RIKEN BioResource Center and a human lung squamous cancer cell line (LK-2) was obtained from the Japanese Collection of Research Bioresources (Osaka, Japan), these 10 cell lines were maintained in RPMI 1640 medium. All mediums were supplemented with 10% foetal bovine serum (FBS; Serum Source International, Charlotte, NC, USA) and 1% penicillin/streptomycin (Nacalai Tesque, Kyoto, Japan). Cultures were maintained at 37 °C in a humidified atmosphere at 5% CO₂. The identity of each cell line was confirmed by DNA fingerprinting via short tandem repeat profiling, as previously described (Ferlay et al, 2015 (link)).