P akt

The human cholangiocarcinoma QBC939 cell line was obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). It was cultured in RPMI 1640 (Sigma, St. Louis, MO, USA) medium supplemented with 10% fetal bovine serum and penicillin and streptomycin (Sigma) at 37°C in a humidified 5% CO₂ atmosphere. Rabbit polyclonal antibodies against FHIT were purchased from Life Technologies (USA). Rabbit polyclonal antibodies against survivin, Bcl-2, caspase-3, AKT and p-AKT were from Genscript Co. (USA).

Human EC Ishikawa cells were obtained from the Chinese Academic of Science cell bank in Shanghai. Medroxyprogesterone acetate (MPA), dimethyl sulfoxide (DMSO) and methylthiazolyldiphenyl-tetrazolium bromide (MTT) were obtained from SIGMA (St. Louis, MO, USA). RPMI 1640 and fetal calf serum (FCS) were obtained from BRL Gibco (Carlsbad, CA, USA). Ethylenediaminetetraacetic acid (EDTA) and sodium carbonate (NaHCO₃) were obtained from Amresco (OH, USA). Annexin-V/propidium iodine apoptosis detection kits were obtained from Bender Med Systems Inc. (Vienna, Austria). Penicillin/streptomycin, Dulbecco's Modified Eagle's Medium (DMEM), L-glutamine, streptomycin and trypsin were obtained from Invitrogen (Pontoise, France). Antibodies against LC3-II/I, PTEN, pAKT, pmTOR, beclin1 and GAPDH were purchased from Genscript Biotechnology (USA).