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.
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.