Human HCC cell lines Bel-7402, Huh7, HepG2, MHCC-97H, and SMMC-7721 and human immortalized normal liver epithelial cell line LO2 were obtained from the Cell Bank of the Shanghai Institutes for Biological Sciences (Shanghai, P.R. China). All HCC cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco). LO2 cells were cultured in RPMI-1640 medium (Gibco) with 10% FBS. Cells were maintained in a humidified incubator containing 5% CO2 at 37°C.
SMMC-7721 and HepG2 cells were plated into 24-well plates and incubated overnight to obtain >70% convergence at the time of transfection. miR-346 mimic, miR-346 inhibitor, and their corresponding control vectors were synthesized from GenePharma (Shanghai, P.R. China). Twenty-nanometer olignonucleotides were transfected into cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s protocol. Small interference RNA of BRMS1 (BRMS1-siRNA) and the negative control (NC) RNA oligonucleotides (GenePharma) were transfected into HepG2 cells using similar methods.
SMMC-7721 and HepG2 cells were plated into 24-well plates and incubated overnight to obtain >70% convergence at the time of transfection. miR-346 mimic, miR-346 inhibitor, and their corresponding control vectors were synthesized from GenePharma (Shanghai, P.R. China). Twenty-nanometer olignonucleotides were transfected into cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s protocol. Small interference RNA of BRMS1 (BRMS1-siRNA) and the negative control (NC) RNA oligonucleotides (GenePharma) were transfected into HepG2 cells using similar methods.