Thle 2

HCC cells (HuH-7 and HCCLM3) and human liver epithelial cell line (THLE-2) were bought from Biovector NTCC (Beijing, China). HCCLM3 and HuH-7 cells were cultured in DMEM (Hyclone, Logan, UT, USA), while THLE-2 cells were grown in BEGM Bullet Kit (Lonza, Walkersville, MD, USA) at 37°C with 5% CO₂. All the mediums were additionally supplemented with 10% FBS (Hyclone) and 1% double antibiotics (Invitrogen, Carlsbad, CA, USA). Cell transfection was carried out using Lipofectamine 3000 (Invitrogen). All oligonucleotides and vectors were synthesized from RiboBio (Guangzhou, China), including circANTXR1 small interference RNA, lentiviral short hairpin RNA and overexpression vector (si-circANTXR1#1/#2/#3, sh-circANTXR1 and oe-circANTXR1) or scrambled controls (si-NC, sh-NC and vector), miR-532-5p mimic and inhibitor (miR-532-5p and anti-miR-532-5p) or scrambled controls (miR-NC and anti-miR-NC), pcDNA XRCC5 overexpression vector (pcDNA-XRCC5) and scrambled control (pcDNA-NC).

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.

Liver epithelial cell-line THLE-2 and HCC cell lines (HuH7, Hep3B, MHCC97-H, LM3) were commercially bought from BioVector NTCC Inc. (Beijing, China). Cell lines were incubated with Roswell Park Memorial Institute-1640 (RPMI-1640; KeyGen, Nanjing, China) containing 10% new-born bovine serum and 1% penicillin-streptomycin. Cells growth was performed in a 5% CO₂ incubator at 37°C.