Mhcc97l

HCC cell lines (BEL7402, SMMC, MHCC97H, MHCC97L, and HuH-7) were purchased from the China Infrastructure of Cell Line Resources. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 1% glutamine. All cells were incubated in a 37°C atmosphere supplemented with 5% CO₂, and passaged once every 2-3 days. The siRNA-RAB27B (siR) and scramble (SCR) lentiviruses containing the gene encoding green fluorescent protein (GFP) were packaged by Gene Pharma Inc. (Shanghai, China). BEL7402 cells were cultured at 8 × 10⁴ cells per well in 6-well plates and infected with siR- RAB27B and SCR lentiviruses, according to the manufacturer’s instructions. PI3K inhibitor LY294002 (sigma, 20 μm) were also added to BEL7402 cells for 24 hours. RAB27B plasmids, containing the complete open reading frame (ORF), were purchased from OriGene (OriGene Technologies, Rockville, MD, USA). Huh-7 cells were added at 10⁵ cells per well in 6-well plates and transfected with RAB27B and Negative Control (NC) plasmids using Lipo3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.