Figure 4
miR-584-3p suppresses the tumorigenesis and aggressiveness of gastric cancer cells via repressing YY1-facilitated MMP-14 expression in vitro. (a and b) Western blot and dual-luciferase assays showing the expression of YY1, MMP-14, VEGF165, and MMP-2 and activity of MMP-14 promoter reporter [pGL3-MMP14 (-1246/ + 199)] in MKN-45 and AGS cells stably transfected with empty vector (mock) or miR-584-3p precursor, and those co-transfected with YY1 (mean ± SD, n = 4). (c) ChIP (using YY1 antibody) and qPCR assay indicating the enrichment of YY1 on MMP-14 promoter in gastric cancer cells stably transfected with mock or miR-584-3p precursor, and those co-transfected with YY1 (mean ± SD, n = 4). (d) Representation (top) and quantification (bottom) of soft agar assay showing the anchorage-independent growth of MKN-45 and AGS cells stably transfected with mock or miR-584-3p precursor, and those co-transfected with YY1 (mean ± SD, n = 5). (e) Representation (top) and quantification (bottom) of matrigel invasion assay indicating the invasion capability of gastric cancer cells stably transfected with mock or miR-584-3p precursor, and those co-transfected with YY1 (mean ± SD, n = 5). (f) Representation (top) and quantification (bottom) of tube formation assay showing the angiogenic capability of gastric cancer cells stably transfected with mock or miR-584-3p precursor, and those co-transfected with YY1 (mean ± SD, n = 5). *P < 0.01 vs. mock + mock.
