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. 2016 Jan 23;7(7):8119–8130. doi: 10.18632/oncotarget.6992

Figure 3. EZH2 knockdown enhances malignant phenotypes in an MDS-derived cell line.

Figure 3

(A) EZH2 mRNA and protein expression was increased in patients with MDS-RCMD and in MDS/leukemia cell lines, but decreased in patients with MDS-RAEB-2. (B) SKM-1 cells with stable overexpression and knockdown of EZH2 were obtained through a lentivirus-mediated transfection system. (C) EZH2 knockdown increased proliferation, and EZH2 overexpression reduced proliferation. (D) EZH2 knockdown reduced cell apoptosis while EZH2 overexpression increased apoptosis. (E) EZH2 knockdown induced an incremental increase in the percentage of cells in S phase, while EZH2 overexpression induced arrest in the G0/G1 phase. (F) EZH2 knockdown increased colony formation. (G) EZH2 knockdown increased cell migration while EZH2 overexpression reduced cell migration. (H) and (I) EZH2 knockdown decreased the expression of adhesion molecules, whereas EZH2 overexpression increased the expression of adhesion molecules. (J) EZH2 knockdown promoted tumor formation in a xenograft model compared to control cells. mRNA and protein analyses identified low EZH2 expression and reduced H3K27me3 levels in xenografts with EZH2 knockdown. (K) Western blot analysis showed that EZH2 knockdown reduced H3K27me3 levels and increased H3K27me1 levels, whereas EZH2 overexpression increased H3K27me3 levels and reduced H3K27me1 levels. H3K27 methylation colorimetric analysis confirmed these results. For cell biological experiments, every assay was carried out three or four times. Representative images are shown. Statistical significance relative to the control group is indicated: *p < 0.05; **p < 0.01; ***p < 0.001 (two-tailed, Student's t-test). Error bars show SEM.