Mk2206

The glioma cell lines U87 and U251 were selected and divided into the Mock group, siNC group, siFAM83D group, FAM83D group, MK2206 group and FAM83D + MK2206 group. Cells in the Mock group received no treatments or transfection; those in the siNC group and siFAM83D group were transfected with negative control siRNA and FAM83D siRNA, respectively. Cells in the FAM83D group were transfected with the pcDNA3.1-FAM83D plasmid, and cells in the MK2206 group were treated with 8 μM AKT inhibitor MK2206 (Beyotime, Jiang Su, China) for 24 h [14] . Cells in the FAM83D + MK2206 group were transfected with the pcDNA3.1-FAM83D plasmid and then treated with MK2206 for 24 h.
Glioma cells in logarithmic growth phase were digested with trypsin, counted and seeded in a 6-well plate at a density of 1 × 10⁶ cells/well for growth, and when the cell confluence reached approximately 70%-80%, transfection was carried out as per the instructions of Lipofectamine^TM 2000 (Invitrogen, USA). Negative control siRNA, FAM83D siRNA and pcDNA3.1-FAM83D plasmids were provided by Shanghai GenePharma Pharmaceutical Technique Co., Ltd. (Shanghai, China).

The HepG2 cell line was provided by China Center for Type Culture Collection. The cells were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin, and cultured in a humidified incubator with 95% air and 5% CO₂ at 37˚C. The cells were subjected to different treatments after they had reached 70% confluence as follows: i) Control group, cells were cultured in DMEM containing 5.5 mM glucose for 24 h; ii) high-glucose group, cells were cultured in DMEM containing 55 mM glucose for 24 h; iii) low-dose oxymatrine treatment group, cells were cultured in DMEM containing 55 mM glucose and 0.1 µM oxymatrine for 24 h; iv) high-dose oxymatrine treatment group, cells were cultured in DMEM containing 55 mM glucose and 1 µM oxymatrine for 24 h; v) metformin treatment group, cells were cultured in DMEM containing 55 mM glucose and 0.1 µM metformin for 24 h; vi) oxymatrine and MK2206 combination treatment group, cells were cultured in DMEM containing 55 mM glucose, 0.1 µM oxymatrine and 3 µM MK-2206 (AKT inhibitor; cat. no. SF2712, Beyotime Institute of Biotechnology) for 24 h. Then, the cells were collected for mRNA and protein expression analysis, or glucose production and uptake assays.