Plc prf 5

Human immortalized liver cell line (HL-7702) and liver cancer cell lines (Huh-7, HepG2, SK-HEP-1, PLC/PRF/5, Hep3B) were purchased from Shanghai Cell Bank (Shanghai, China). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA), 1% penicillin–streptomycin (Sangon, Shanghai, China), and 2 mM glutamine (Invitrogen, Carlsbad, CA, USA) at 37 °C in a 5% CO₂ humidified atmosphere.

Twenty paired fresh-frozen HCC and para-tumor tissue samples were used in quantitative real-time PCR (qRT-PCR) analysis. HCC and para-tumor tissue samples were surgically resected from HCC patients who underwent hepatectomy at the Peking Union Medical College Hospital (PUMCH, Beijing, China) between 2010 and 2014. All diagnoses were confirmed by pathology. This study was approved by the Ethics Committee of PUMCH, and informed consent was obtained from each patient.
The 293T cell line and HCC cell lines HepG2, SMMC-7721, PLC/PRF/5, Huh7 and SK-hep-1 were obtained from Shanghai Cell Bank, Chinese Academy of Sciences, and cultured as recommended by the supplier. Human diploid fibroblast cells 2BS, IMR90 and MRC-5 were obtained from National Institute of Biological Products, Beijing, China, and cultured as recommended by the supplier. Cells were transiently transfected with plasmids using Lipofectamine 2000 Reagent (Invitrogen), and siRNAs were transfected using Lipofectamine RNAiMAX Reagent (Invitrogen, Carlsbad, USA) following the manufacturer’s protocol. Forty-eight hours after transfection, cells were harvested and lysed to evaluate the transfection efficiency.