Hsc 4

OSCC cell lines (PECAPJ41 and HSC-4) and normal oral cells (HOK) were purchased from Nanjing Kebai Biotechnology Co., Ltd (Nanjing, China). PECAPJ41 and HSC-4 cells were cultured with RPMI 1640 (Thermo Fisher Scientific, USA) containing 10% fetal bovine serum (FBS). HOK was cultured in MEM (Thermo Fisher Scientific, USA) containing 10% FBS. All cells were cultured in a constant temperature and humidity box at 37°C and 5% CO₂. siRNA negative control (si-control), GACAT1-siRNA, inhibitor NC and miR-149 inhibitors were constructed by Geneseed Co., Ltd. (Guangzhou, China). Cells were cultured in 96 well plates 24 hours before transfection. According to the manufacturer’s protocol, Lipofectamine 2000 (Invitrogen, CA) was used for transfection.

KOSC-2 cl3-43 (KOSC-2), HSC-2, HSC-3, HSC-4, and SCC-4 human oral cancer cell lines were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). KOSC-2 cells were grown in RPMI medium (Thermo Fisher Scientific, Waltham, MA, USA). HSC-2, HSC-3, and HSC-4 cells were grown in MEM (Thermo Fisher Scientific). SCC-4 cells were grown in Dulbecco’s Modified Eagle Medium (Thermo Fisher Scientific). All media were supplemented with 10% fetal bovine serum (Thermo Fisher Scientific).

Five OSCC cell lines, HSC-3, HSC-4, Cal-27, Cal-33 and SCC25 were used in this research. HSC-3 and HSC-4 were purchased from the cell bank of Japanese Collection of Research Bioresource (JCRB, Shinjuku, Japan). Cal-27 and SCC25 were purchased from American Type Culture Collection (ATCC, Manassas, USA). Cal-33 was obtained from the European Collection of Cell Cultures (ECACC). HSC-3 and HSC-4 cells were cultured in DMEM (Gibco, Eggenstein, Germany) supplemented with 10% fetal bovine serum (FBS; Gibco), and Cal-27, Cal-33 and SCC25 cells were maintained in DMEM/Ham's F-12 (1:1) supplemented with 10% FBS and hydrocortisone (400 ng/ml) (Sigma-Aldrich, Poole, UK) under 5% CO2 and humidified air atmosphere at 37°C.