Hepg2

Human HCC cell lines, BEL-7404, BEL-7402, HepG2, and SMMC-7721, were obtained from Shanghai Genechem Co., Ltd. (China). Cells were maintained in Dulbecco's Modified Eagle Medium (DMEM; Corning, NY, USA) plus 10% fetal bovine serum (Bovogen, Melbourne, Australia) in a 5% CO2 incubator at 37.0°C.

Human hepatic cell line HepG2 was obtained from GeneChem (Shanghai, China). STR profiling and mycoplasma contamination testing were provided by GeneChem. HepG2 cells were cultured in Dulbecco’s modified Eagle’s medium (HyClone, Logan, UT, USA) supplemented with 10% fetal bovine serum (Biological Industries, Kibbutz Beit Haemek, Israel). Cells were cultured in a humidified 5% CO₂ incubator at 37°C. Exo-clear complete cell growth medium (System Biosciences, Palo Alto, CA, USA) was used for extraction of exosomes from cell culture supernatant.

Human LC cell lines (HepG2 and SNU-182) and a transformed human liver epithelial-3 cell line (THLE-3) were purchased from American Type Culture Collection. All cell lines were authenticated via STR profiling. Cells were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO₂. Specific small interfering (si)RNAs targeted against TPT1-AS1 were used, including si-TPT1-AS1#1 (5′-AAGGTACCGAAAGCACAGTAA-3′), si-TPT1-AS1#2 (5′-AACCATCACCTGCAGGAAACA-3′) and a scrambled siRNA control (si-NC) (5′-AACCATCACTTACAAGAAACC-3′), which were purchased from Shanghai GeneChem Co., Ltd. HepG2 and SNU-182 cells (2×10⁴ cells/well) were transfected with 50 nM si-TPT1-AS1#1, si-TPT1-AS1#2 or si-NC using Lipofectamine^® 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) for 48 h at 37°C. At 48 h post-transfection, cells were used for subsequent experiments.

The human liver cancer cell lines HepG2, SMMC7721, BEL-7404, SK-hep-1 and Huh7 were purchased from Shanghai Genechem Co., Ltd. (Shanghai, China). HCC-LM3, which is a highly invasive hepatocellular carcinoma cell line, was provided as a gift from the laboratory of the Eastern Hepatobiliary Surgery Hospital affiliated to the Second Military Medical University. HCC-LM3 cell line was approved by the Institute Ethics Committee at the Affiliated Hospital of Nantong University. The HCC-LM3, HepG2 and Huh7 liver cancer cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco) at 37°C under 5% CO₂. The liver cancer cell lines SMMC7721 and BEL-7404 were cultured in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) medium with 10% FBS (Gibco) at 37°C under 5% CO₂. The SK-hep-1 liver cancer cell line was cultured in MEM (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Gibco) at 37°C under 5% CO₂.

Cancer cell lines (MCF7, MGC803, EC109, HepG-2, PC-3, A549, OC-314, KYSE-450 and SK-N-SH) were purchased from GeneChem (Shanghai, China), cancer cell lines (OVCAR-8, SKOV3 and Caov-3) were purchased from Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cells were cultured in RPMI 1640 (Hyclone, Logan, UT, USA), supplemented with 10% foetal bovine serum (Hyclone, Logan, UT, USA) in a humidified CO₂ (5%) incubator at 37°C.