Recombinant human il rhil 2

HCC cell line SMMC7721, HepG2, human NK cell line NK-92, and 293T cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). SMMC7721, HepG2, and 293T cells were maintained in DMEM with 10% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and 1% penicillin/streptomycin. NK-92 cells were maintained in α-minimum essential medium (Invitrogen, Carlsbad, CA, USA) supplemented with 12.5% FBS (Sigma-Aldrich), 12.5% horse serum (Gibco, Grand Island, NY, USA), 100 U/mL recombinant human IL (rhIL)-2 (PeproTech, Rocky Hill, NJ, USA), and 0.1 mM β-mercaptoethanol. The isolated primary NK cells were cultured in complete RPMI medium (Gibco) supplemented with 10% heat-inactivated FBS (Sigma-Aldrich), 2 mM L-glutamine, and 100 U/mL rhIL-2 (PeproTech). All cells were incubated at 37℃ in an incubator of 5% CO₂.
miR-506 mimic (miR-506), miRNA negative control (miR-NC), miR-506 inhibitor (anti-miR-506), inhibitor control (anti-miR-NC), pcDNA-STAT3, pcDNA empty control (pcDNA), siRNA specially against STAT3 (si-STAT3), and siRNA control (si-NC) were purchased from GenePharma (Shanghai, China). NK-92 and primary NK cells were seeded into 96-well plates and transfected with the abovementioned nucleotides or plasmids, using Lipofectamine 2000 (Invitrogen). Co-culturing or RNA extraction was performed 48 hours after transfection.