Fig. 6

MLL3 elevates GRHL2 to restrict GC cell proliferation, migration, invasion, and vessel-like tube formation of HUEVCs. a Correlation between MLL3 and GRHL2 expression in GC obtained by GEPIA (p = 4.4e-16). b RT-qPCR was used to detect the expression of GRHL2 in GC tissues. * p < 0.05 vs. Adjacent normal tissues. c The effect of MLL3 on GRHL2 expression in NUGC-3 and HGC27 cells was detected by RT-qPCR. * p < 0.05 vs. GES-1 cells; # p < 0.05 vs. oe-NC-transfected cells (NUGC-3); & p < 0.05 vs. oe-NC-transfected cells (HGC27). d ChIP assay to detect MLL3, H3K4me1 and H3K27ac enrichment in GRHL2 gene enhancer region. * p < 0.05 vs. the IgG group; # p < 0.05 vs. the oe-NC group. e RT-qPCR was used to detect the expression of MLL3 and GRHL2 in each group of cells. f CCK-8 assay was adopted to detect the viability of cells. g TUNEL assay was used to detect apoptosis of cells (200 ×). h Transwell assay was utilized to detect the migration of cells (200 ×). i Transwell assay was conducted to detect the invasion of cells (200 ×). j Matrigel-based angiogenic assays were performed to detect numbers of vessel-like tubes formed in vitro (100 ×). k, l Western blot assay was used to detect the expression of proteins normalized to GAPDH in each group. e-l * p < 0.05 vs. sh-NC-transfected cells; # p < 0.05 vs. sh-MLL3 + oe-NC-transfected cells. Unpaired t test was used for the comparison between the two groups, and two-way ANOVA and Bonferroni test were performed at different time points