Hcclm3

The cell lines (L02, Huh7, HCCLM3 and Hep3B) were obtained from Procell (Wuhan, China), and maintained in Dulbecco's modified Eagle's medium (DMEM) with fetal bovine serum (FBS) (10%) and penicillin/streptomycin (100 U/mL, Gibco, Grand Island, NY, USA) under 5% carbon dioxide (CO₂) at 37 °C.

HCC cell lines (HCCLM3 and Huh7) and human liver epithelial cell line (THLE-2) were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China). HCCLM3 and Huh7 cells were grown in Dulbecco’s modified Eagle medium (DMEM; Invitrogen, Carlsbad, CA, USA), and THLE-2 cells were cultured in BEGM Bullet Kit (Lonza, Walkersville, MD, USA). In addition, all the media should be supplemented with 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and 1% penicillin/streptomycin (Invitrogen). All cells were incubated at 37°C with 5% CO₂.
Circ_0046599 small interference RNA and overexpression plasmid (si-circ_0046599#1/#2 and circ_0046599) or negative controls (si-NC and circ-NC), miR-1258 mimic and inhibitor (miR-1258 and anti-miR-1258) or negative controls (NC and anti-NC), Ribophorin II (RPN2) overexpression plasmid (RPN2) and negative control (vector) were synthesized by GenePharma (Shanghai, China). Lentiviral short hairpin RNA against circ_0046599 (sh-circ_0046599) and negative control (sh-NC) were obtained from RiboBio (Guangzhou, China). Lipofectamine 3000 (Invitrogen) was used to transfect the above plasmids and oligonucleotides into cells.