Bgc 823

Human gastric normal epithelium cell line (GES-1) and GC cells (BGC-823, SGC-7901, and MGC-803) were purchased from Shanghai Institutes for Biological Science, China. BGC-823, SGC-7901 and MGC-803 cells were cultured in RPMI 1640 medium (KeyGene, Nanjing, China). GES-1 cells were cultured in DMEM medium (KeyGene, Nanjing, China) supplemented with 1% penicillin/streptomycin, l-glutamine, and 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA). Culture plates were incubated at 37°C in a humidified atmosphere with 5% CO².
The siRNA specifically targeting RP11-138J23.1, HuR and VAV3 was designed and synthesized by Invitrogen (Invitrogen, USA). The nucleotide sequences of all siRNAs were shown in Supplementary Table S1. The cells were transfected with siRNA using RNAiMAX (Invitrogen, USA). In addition, RP11-138J23.1 overexpression plasmid and vectors containing shRNA targeting RP11-138J23.1 was constructed and transfected with XtremeGENE HP DNA transfection reagent (Roche, Basel, Switzerland).

Luteolin is purified and provided from Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine (Beijing, China), and the purity of the product was over 98%, detected by HPLC (UV). Gastric tumor cell lines of SGC7901, SGC7901/DDP, HGC27, MGC803, BGC803 and BGC823 were purchased from Keygene (Jiangsu, China). Monoclonal antibodies against HSP-90, STAT3 and phosphor-STAT3 (Tyr705), STAT1 and phosphor-STAT1 (Tyr701), Akt, phosphor-Akt (Ser473), Erk and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) were purchased from Cell Signaling Technology (Boston, MA, USA). Monoclonal antibodies against SHP-1, SHP-2, Bcl-xl, Survivin, Mcl-1, SOCS3, PIAS3, gp130, ki0–67, activated caspase-3 and GAPDH were purchased from Abcam (Burlingame, CA, USA). BCA Protein Assay Kit was purchased from Pierce (Rockford, IL, USA). l-Ascorbic acid was purchased from Cayman (Ann Arbor, MI, USA), α-Tocopherol was purchased from Selleckchem (Houston, TX, USA). DCFDA was purchased from Sigma-Aldrich (St. Louis, MO, USA).