Kyse170

The esophageal cancer cell lines Eca109, TE1, KYSE30, YES2, EC9706, KYSE170 and KYSE150 were obtained from Procell Life Science&Technology Co. (Wuhan, China). Eca109, YES2, EC9706, KYSE170 and KYSE150 were cultured in DMEM medium (GIBCO, USA) containing 10% fetal bovine serum (FBS; Biological Industries, Beit-Haemek, Israel) and 1% penicillin-streptomycin (Solarbio, China) with 5% CO₂ at 37 °C in a humidified incubator. TE1 and KYSE30 cells were cultured in 1640 medium (GIBCO, USA) containing 10% FBS and 1% penicillin-streptomycin with 5% CO₂ at 37 °C in a humidified incubator.

The human ESCC cell lines TE‐1 (RRID:CVCL_1759), KYSE‐30 (RRID:CVCL_1351), KYSE‐150 (RRID:CVCL_1348), and KYSE‐170 (RRID:CVCL_1358) were purchased from Procell Life Science & Technology Co., Ltd (Wuhan, China) and stored in our laboratory. These cell lines have been authenticated in the past 3 years using short tandem repeat analysis. TE‐1, KYSE‐30, and KYSE‐150 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium, whereas KYSE‐170 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% heat‐inactivated FBS (GIBCO, Thornton, NSW, Australia) in a 37 °C sterile incubator with 5% CO₂. In this study, the cells used in all experiments were mycoplasma‐free.

Four kinds of human esophageal cancer cells inculding TE1, KYSE30, KYSE150 and KYSE170 were chosen to study, which were purchased from Procell Life Science&Technology Co., Ltd. (Wuhan, Hubei, China). There are no mycoplasma contamination among them. All cell lines were culture with RPMI1640 medium (GIBCO, USA) containing with 10% fetal bovine serum (FBS), 100 U/ml of penicillin and 100 μg/ml streptomycin, which were maintained at 37 °C in a humidified atmosphere of 5% CO₂. For drug treatment, cells were cultured with or without 10 ng/mL recombinant human RANTES (CCL5) (Peprotech, USA) in medium. For transfection, siRNAs were supplied by Guangzhou RiboBio Co.,Lid.( Guangzhou, China). The hsa_circ_0060927 sequence was cloned into the pLC5-ciR vector and synthesized by Geneseed Biotech Co.( Guangzhou, China). According to the manufacturer’s instruction, ESCC cells inoculated in 6-well cell plates (1 × 10⁶) were transfected with pLC5-ciR vector or siRNA by using FuGENE HD transfection Reagent (Promega, USA) or HiperFect transfection Reagent (Promega, USA). 6 h later, the complete medium was used to instead of the transfection medium. After a further culture for 48 h, the cells were collected.