Figure 2. EED inhibits miR-338-5p expression by promoting methylation. (A) Absolute miR-338-5p expression in profile GSE23739 obtained from GEO database (https://www.ncbi.nlm.nih.gov/gds) analyzed by R language. The blue box on the left represents the expression of normal samples, and the red box on the right represents the expression of GC samples (p = 5.772E-03). (B) RT-qPCR determination of miR-338-5p expression in GC tissues and adjacent normal tissues (n = 97). (C) miR-338-5p expression in GC tissues was negatively correlated with EED expression in GC tissues (n = 97). (D) Evaluation of EED knockdown efficiency in MGC-803, and HGC-27 cells. (E) ChIP assay to assess enrichment of EED in the miR-338-5p promoter. (F) Enrichment of H3K27me3 in the miR-338-5p promoter. (G) RT-qPCR to examine EED and miR-338-5p expression after overexpressing EED and DZNep treatment, with β-actin and U6 as internal control, respectively. Measurement data are expressed as mean ± standard deviation. * p < 0.05. GC tissues were compared with adjacent normal tissues by paired t test, and other comparison between two groups was analyzed using an unpaired t test. Data comparison among multiple groups was performed using one-way ANOVA with Tukey's post hoc test. Pearson’s correlation analysis was carried out for the correlation between miR-338-5p and EED expression in GC tissues. Cell experiments were repeated 3 times independently.