Bcpap

Thyroid cancer cell line BHT101 and BCPAP was purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). BHT101 cell was grown in DMEM medium (Gibco) containing 20% heat‐inactivated FCS (Gibco) and glutamine; BCPAP cells was maintained in RPMI 1640 medium containing 10% fetal bovine serum (Gibco) and 2 mmol/L l‐glutamine (Invitrogen, Carlsbad, CA) at 37°C with 5% CO₂. The LINC00704 and negative control siRNAs (Invitrogen, Carlsbad, CA) were transfected into BHT101 and BCPAP cells using RNAiMAX (Invitrogen) according to the manufacturer's protocol. Forty‐eight hours after transfection, the cells were harvested for further experiment. The LINC00704 siRNA sequences are siRNA 1#, 5′‐GCUUGCUCUCACAGCCAUUTT‐3′ siRNA 2#, 5′‐GGAACCUCCUUUCUUACAUTT‐3′.

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.