Fig. 4

miR-598-3p directly targets RMP and IGF1r. A Heatmap showing differentially expressed genes in MKN45 cells with miR-598-3p overexpression (miR-598-3p) or knockdown (Anti miR-598-3p). B, C Volcano plot of miR-598-3p overexpression (B) or knockdown (C) versus control based on the RNA-seq analysis. D Venn diagram showing differentially expressed genes and prognostic genes derived from the indicated gene sets: miR-598-3p down (decreased gene sets in group of miR-598-3p in A), Anti-miR-598-3p up (increased gene sets in group of Anti-miR-598-3p in A), targeted genes predicted by miRpathDB or Targetscan 7.1. E, F Gene set enrichment analysis (GSEA) of the RNA-seq data set in A. G, H TargetScan 7.1 predicted the miR-598-3p binding sites in RMP and the IGF1r 3ʹUTR, as well as the designed mutant 3ʹUTR. I, J Luciferase reporter assay of the 3ʹUTR of RMP (I) or IGF1r (J) in HEK-293T cells cotransfected with the indicated plasmid. K–N mRNA expression of IGF1r and RMP in SGC-7901 or MKN45 cells that were cotransfected with the indicated plasmid, after which the cells were treated with or without hypoxia. O, P Luciferase reporter assay in HEK-293T cells cultured in hypoxia and transfected with wild-type (WT) or mutant (MUT) 3ʹUTR of RMP (O) and IGF1r (P). Q–S Real-time PCR analysis of miR-598-3p (Q), RMP (R) and IGF1r (S) levels in tumor tissue and relative paratumor tissue of 20 gastric cancer patients