Kyse150

Human ESCC cell lines (EC109, EC9706, KYSE150 and TE-1) and HEK-293T were obtained from the Hunan Fenghui Biotechnology Co., Ltd. (Changsha, China). These cells were cultured in high glucose DMEM medium (C11995500, Gibco, NY, USA) supplemented with 10% fetal bovine serum (04-001-1A, FBS, Biological Industries, Israel) and penicillin–streptomycin solution (100 U/ml, P1400, Solarbio, Beijing, China). Melatonin (#14427, Cayman Chemical Company, Michigan, USA), SAHA (HY-10221, MedChemExpress, USA), Lf3 (HY-101486, MedChemExpress, USA) and 10058-F4 (HY-12702, MedChemExpress, USA) stock solution was prepared in DMSO and diluted in culture media immediately prior to the subsequent experiments. The ESCC cells were treated with these chemical compounds for 48 h and then harvested for further analyses.

The human esophageal cancer cell line Eca109 and KYSE150 cells were purchased from Hunan Fenghui Biotechnology Co., Ltd (CL0092; CL0493, Changsha, Hunan, China) and cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) at 37°C in a 5% CO₂ atmosphere. The over-circRNA TCFL5, sh-circRNA TCFL5, miR-543 mimics, miR-543 inhibitor, over-FMNL2, and sh-FMNL2 were provided by GenePharma (Shanghai, China). Transfections of 50 nM miR-543 mimics, 100 nM miR-543 inhibitor, 50 nM sh-RNA, and 2 μg RNA overexpressing vector were performed by using the Lipofectamine 3000 transfection reagent (Invitrogen) following the manufacturer's instruction when the confluence reached 60-90%. The function of these factors on cell proliferation, migration, and invasion was analyzed at 48 h after transfection.