Mir 18a inhibitor

Lipofectamine 2000 (Invitrogen) was used for oligonucleotide transfection in this study per the manufacturer’s recommendations. The miR-18a mimic, miR-18a inhibitor, and negative control (NC) miRNA were designed and synthesized by RiboBio (Guangzhou, P.R. China). IRF2 or CBX7 expression constructs (1 μg; OriGene, Beijing, P.R. China) were transfected into cells with or without miR-18a mimic. After transfection for 48 h, the cells were collected and analyzed by quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot to evaluate the transfection efficacy.

Human HCC cell lines HepG2 and SMMC7721 and normal liver cell line LO2 were purchased commercially from Tianjin Medical University Cancer Institute and Hospital. DMEM or RPMI-1640 media (Corning Incorporated, Corning, NY, USA) containing 10% FBS (Hyclone, Logan, UT, USA) were used to maintain cells. Cells and media were kept in a 5% CO₂ environment in a incubator at 37°C. The medium was replaced every 2 days. miR-18a mimic (Ribobio, Guangzhou, China), miR-18a inhibitor (Ribobio), and Bcl2L10-overexpressing plasmids (Genechem, Shanghai, China) were used to transfect HCC cell lines in order to regulate miR-18a and Bcl2L10 expressions.