Ec9706

Six human ESCC cell lines (TE-1, TE-8, KYSE150, KYSE450, Eca-109 and EC9706) and the normal human esophageal epithelial cell line HEECs (cat. no. BNCC337729) were purchased from the Shanghai Institute of the Chinese Academy of Sciences (Shanghai, China). Another normal human esophageal epithelial cell line Het-1A was purchased from American Type Culture Collection (Manassas, VA, USA). TE-1, TE-8, Eca-109, EC9706 and Het-1A cells were cultured in Roswell Park Memorial Institute medium-1640 (Thermo Fisher Scientific Inc., Waltham, MA, USA), and KYSE150, KYSE450 and HEECs cells were cultured in Dulbecco's modified Eagle's medium (Thermo Fisher Scientific, Inc.). These media were supplemented with 10% fetal bovine serum (Invitrogen; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin in a humidified atmosphere at 37°C with 5% CO₂.

The human normal esophageal cell line, Het-1A (BNCC337688), and EC cell lines [TE-1 (BNCC100151), Eca-109 (BNCC337687) and EC9706 (BNCC339892)] were obtained from Bena Culture Collection (BNCC, Beijing, China). The Het-1A, Eca-109 and EC9706 cell lines were grown in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc.) containing 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin. The TE-1 cell lines were cultured in RPMI-1640 medium containing 10% FBS. All of the cells were placed in a wet incubator with 5% CO₂ at 37°C.

Human ESCC cell lines (EC9706, EC109, KYSE30 and KYSE150) and a human esophageal epithelial cell line (Het-1A) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). These cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium containing 10% fetal bovine serum (FBS), 10,000 units/mL of penicillin and 10,000 g/mL of streptomycin. The culture conditions were 37°C and 5% CO₂. Cells were harvested during logarithmic growth phase, and seeded in 6-well plates at 1×10⁵ cells/well. The transfection products, including si-MAFG-AS1 (5′-GGGCAAUUCCAACCAAGAAAC-3′), si-NC (negative control siRNA; 5′-AAUUCUCCGAACGUGUCUCGUGU-3′),6 (link) miR-765 mimics and control mimics were all synthesized by GenePharma (Shanghai, China). EC109 and EC9706 cells were transfected with 100 ng of the vector or 50 nM of the indicated oligonucleotide by Lipofectamine™ 2000 (Invitrogen, CA, USA). For overexpression, MAFG-AS1 was cloned into mammalian expression vector pcDNA3.1 (Invitrogen, CA, USA) to construct overexpressing plasmid p-MAFG-AS1. Vector pcDNA3.1 was used as a vector control. After 48 h of transfection, cells were used for the next detection.