293 t cells

Starbase3.0 (http://starbase.sysu.edu.cn) was used to predict the binding between miR-152-3p and circRERE or CD47. To analyze the interaction between miR-152-3p and circRERE, the circRERE luciferase plasmids of wild-type (circRERE wt) and mutant-type (circRERE mut) were generated by the molecular cloning into the pmirGLO plasmid (Promega, Madison, WI, USA). For miR-152-3p and CD47, the luciferase plasmids CD47 3′UTR wt and CD47 3′UTR mut were constructed. These plasmids were respectively co-transfected with mimic NC or miR-152-3p mimic into 293 T cells (BioVector NTCC Inc.) for 48 h, then the luciferase activity examination was carried out through the Luc-Pair™ Duo-Luciferase HS Assay Kit (GeneCopoeia, Rockville, MA, USA).

The normal human hepatocyte THLE-2, HCC-derived cell lines (HepG2 and Huh7) and 293T cells were purchased from BioVector NTCC Inc. (Beijing, China). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Grand Island, NY, USA) harboring with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin/streptomycin at 37°C with 5% CO₂.
The miRNA mimic or inhibitor targeting miR-296-5p (miR-296-5p mimic or miR-296-5p inhibitor) and their corresponding negative control (miRNA NC or inhibitor NC) were purchased from RIBOBIO (Guangzhou, China). Short hairpin RNA (shRNA) targeting NEAT1 (sh-NEAT1), shRNA scramble control (sh-NC), small interfering RNA (siRNA)targeting NEAT1 (si-NEAT1), siRNA negative control (si-NC), pcDNA3.1-CNN2 overexpression vector (pcDNA-CNN2), pcDNA3.1 empty vector (pcDNA-Control) were synthesizedby Genepharma (Shanghai, China). Subsequently, Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA) was used to transfect these oligonucleotides or vectors into HepG2 and Huh7 cells following the protocol of the manufacturer.