Eif4a3 sirna

circSMAD2 siRNAs (circSMAD2 siRNA1, circSMAD2 siRNA2 and circSMAD2 siRNA3), eiF4A3 siRNA or negative control RNAs (all at 10 nM) were transfected into GBC cells using Lipofectamine 2000 (Invitrogen, Carlsbad, CA). circSMAD2 siRNAs, eiF4A3 siRNA and negative control RNAs were purchased from GenePharma (Shanghai, China).
For SMAD2 overexpression, GBC cells were transfected with pcDNA3.1 or pcDNA3.1-SMAD2 (SMAD2 oE) by using Lipofectamine 2000 (Invitrogen). pcDNA3.1 and pcDNA3.1-SMAD2 were obtained from Genepharma.

The circ_0003159 overexpression plasmid, eIF4A3 overexpression plasmid, and the empty plasmid (vector or NC) were purchased from YRBIO (Changsha, China). eIF4A3 siRNA (si-eIF4A3, 5′-AGACAUGACUAAAGUGGAA-3′) and the negative control sequences were designed and synthesized by GenePharma (Shanghai, China). The GC cells were transfected with the plasmid or siRNA by the Lipofectamine 3000 reagent (Invitrogen, CA, USA). After 24 h later, ICA was added into the GC cell culture and culture together for 48 h.

Four GC cell lines, AGS, BGC-823, SGC-7901, and HGC-27, and the human immortalized normal gastric epithelial cell line GES-1 were provided and characterized by GeneChem Co., Ltd. (Shanghai, China). Hsa_circ_100290 and EIF4A3 siRNA and the corresponding negative control sequences were designed and synthesized by GenePharma (China). The si-circRNA_100290 sequence of the sense strand was 5’-CUCAUGCUUAGGCUUGAUUdTdT-3’; the sequence of the antisense strand was 3’-dTdTGAGUACGAAUCCGAACUAA-5’. The si-EIF4A3 sequence of the sense strand was 5’-CGAGCAAUCAAGCAGAUCAdTdT-3’, and the sequence of the antisense strand was 3’-dTdTGCUCGUUAGUUCGUCUAGUdTdT-5’. AGS and HGC-27 cells were prepared for si-circRNA_100290 and si-EIF4A3 transfection using Lipofectamine reagent (GenePharma, China) according to the manufacturer’s protocol. The knockdown efficiency was detected by quantitative real-time polymerase chain reaction (qRT-PCR) 48 h after transfection.