Fig. 6

ESR1 is a downstream target gene of miR-204-5p. (a) Downstream target genes of miR-204-5p were predicted using starBase, miRcode, TargetScan, miRDB, and miRWalk. (b) RT-qPCR was used to analyze the mRNA expression levels of IGFBP5, NOTCH2, and ESR1 in human GCs. (c) Bioinformatics software was used to predict the binding sites of miR-204-5p and ESR1. (d) Dual-luciferase reporter assay was performed to verify the target-binding relationship between miR-204-5p and ESR1. (e–f) Expression levels of ESR1 mRNA and protein following transfection with miR-204-5p mimics or inhibitors were detected using RT-qPCR and western blotting, respectively. ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001