Kyse30

Four ESCC cell lines (EC109, EC9706, KYSE30, and KYSE450) were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). A normal esophageal epithelial cell (Het-1A) was purchased from Jenniobio Biotechnology (Guangzhou, China). All cells were cultured in RPMI-1640 medium (Hyclone, USA) supplemented with 10% fetal bovine serum (10% FBS), and maintained in a humidified incubator at 37°C with 5% CO_2.

Human ESCC cell lines KYSE30, KYSE150, KYSE450, KYSE510, and TE-1 purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) were cultured in RPMI 1640 medium (Gibco, USA) containing 10% fetal bovine serum (FBS; Gibco, USA) as previously described [42 (link)]. The immortalized human esophageal epithelial cell line Het-1A and human embryonic kidney 293T cells, obtained from the ATCC (Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium culture medium supplemented with 10% FBS. Cells were kept in a humidified incubator with 5% CO₂ at 37 °C. All cells were periodically authenticated by using the short tandem repeat analysis and tested for mycoplasma contamination by PCR.

Human esophageal cancer cell lines (Eca109, Eca9706, KYSE30, KYSE510, KYSE520, KYSE140 and KYSE150) were obtained either from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) or purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany) and cultured at 37°C with 5% CO2 in Dulbecco’s Modified Essential Medium (DMEM) medium (Hyclone, Logan, Utah, USA) supplemented with 10% fetal bovine serum (FBS) (Gibico, Carlsbad, California, USA). Human esophageal epithelial squamous cell NE1 was a kind gift from Dr. Yang XZ at Cancer Hospital and Institute of Guangzhou Medical University (Guangzhou, China) were cultured in a 1:1 mixture of defined keratinocyte serum free medium with growth supplements and EpiLife medium with 60 μM Calcium (Invitrogen,Carlsbad, California, USA). BAY 11-7082 and TNFα were purchased from Selleck (Huston, Texas, USA) and dissolved according to the manufacturer instructions. The fluorescent stain 4',6-diamidino-2-phenylindole (DAPI) and second antibodies used for immunofluorescence were purchased from Invitrogen (Carlsbad, California, USA).

Human normal esophageal epithelial cell line HEEC and ESCA cell lines Eca109, TE-1, EC9706, EC8712 and KYSE30 cells were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were maintained in DMEM medium plus 10% fetal bovine serum in humidified 5% CO₂ at 37°C.