Tpc 1

Papillary thyroid cancer cell lines TPC-1 and BCPAP cells were purchased from the National Infrastructure of Cell Line Resource (Beijing, China). Nthy-ori-3-1 cells was kindly provided by Professor. Yang Yan. TPC-1, BCPAP, and Nthy-ori-3-1 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Cleveland, TN, USA), with 10% fetal bovine serum (FBS) (Gibco, Cleveland, TN) and 1% penicillin/streptomycin in a 37 °C/5% CO2 incubator. Lipofectamine 2000 (Invitrogen, USA) was used to transfect small interfering RNA into TPC-1 cells. Forty-eight hours after transfection, the cells were collected and analyzed by western blot. The small interfering RNA was synthetized in Sangon Biotech (Shanghai, China). The small interfering RNA sequences were: siNC:UUC UCC GAA CGU GUC ACG UTT; siSGLT2 1#:CGACAAAUACCUGGGAGCAAUTT; si SGLT2 2#:ACCAUGAUUUACACGGUGACATT.

The human PTC cell line, BCPAP, the German Collection of Micro-organisms and Cell Cultures (Braunschweig, Germany). The human PTC cell line, TPC1 and normal thyroid cell line Nthy-ori-3-1were obtained from Chinese Science Institute (Shanghai, China), and the human PTC cell line, K1, were purchased from the European Collection of Authenticated Cell Cultures (ECACC, U.K.). The human PTC cell line, IHH4, was obtained from Health Science Research Resources Bank (Osaka, Japan). Cell culture medium was prepared according to the suppliers’ instructions. The BCPAP and Nthy-ori-3-1 were cells were cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, U.S.A.) supplemented with 10% FBS (Gibco, Carlsbad, CA, U.S.A.). The TPC1 and K1 were maintained in DMEM high glucose (Gibco, Carlsbad, CA, U.S.A.) supplemented with 10% FBS. IHH4 was maintained in a mixture (1:1) of RPMI 1640 and DMEM high glucose medium supplemented with 10% FBS. All cells were cultured at 37°C, in a humidified atmosphere with 5% CO₂.
The plasmids of pcDNA3.1 (control) and pcDNA3.1-GAS8-AS1, GAS8-AS1 siRNA (si-GAS8-AS1) and scramble, miR-135b-5p mimic and negative control (NC), CCNG siRNA (si-CCNG), and scramble were obtained from GenePharma (Shanghai, China). TPC1 and BCPAP were transfected using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s protocol.

The thyroid cancer cell lines BC-PAP, TPC-1, 8505c and FTC133, and the normal thyroid cell line NTHY-ORI3-1 were purchased from Gibco (Thermo, Waltham, MA, USA). The thyroid cancer cell lines BC-PAP, TPC1, and 8505c were cultured with Dulbecco’s modified Eagle’s medium (DMEM) (Thermo, Gibco, MA, USA), and supplemented with 10% fetal bovine serum (FBS) (Thermo, Gibco, MA, USA) and 1% penicillin–streptomycin (Thermo, Gibco, MA, USA). The thyroid cancer cell line FTC133 and normal thyroid cell line NTHY-ORI3-1 were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium containing 10% FBS (Thermo, Gibco, MA, USA) and 1% penicillin–streptomycin (Thermo, Gibco, MA, USA). All cells were cultured at 95% humidity and 5% CO₂ at 37 °C.