MiR-451 inhibits proliferation of esophageal carcinoma cell line EC9706 by targeting CDKN2D and MAP3K1

Figure 2 CDKN2D and MAP3K1 overexpression reverses the effect of miR-451. A: CDKN2D and MAP3K1 protein levels were detected by Western blot assay. Western blot assay showed that transfection of miR-451 mimics inhibited the expression of CDKN2D or MAP3K1. Co-transfection of pcDNA3.1-CDKN2D or pcDNA3.1-MAP3K1and miR-451 abrogated the effects of miR-451 on CDKN2D or MAP3K1 expression. β-actin was used as a reference; B: The expression of CDKN2D could partially reverse the anti-proliferation function of miR-451. Colony formation assays were performed. ^aP < 0.05 vs control group; C: The expression of MAP3K1 could partially reverse the anti-proliferation function of miR-451. Colony formation assays were performed. ^aP < 0.05 vs control group; D: The expression of CDKN2D did not reverse the pro-apoptotic function of miR-451. Cells were transfected with pcDNA3.1-CDKN2D (not including 3’-UTR) or (and) miR-451. The cell apoptosis was assessed using flow cytometry assay; E: The expression of MAP3K1 reversed the pro-apoptotic function of miR-451. Cells were transfected with pcDNA3.1-MAP3K1 (not including 3’-UTR) or (and) miR-451. The cell apoptosis was assessed using flow cytometry assay; F: The expression of CDKN2D reversed the anti-migration function of miR-451. Cells were transfected with pcDNA3.1-CDKN2D (not including 3’-UTR) or (and) miR-451. The cell invasion was assessed using transwell assay; G: The expression of MAP3K1 reversed the anti-migration function of miR-451. Cells were transfected with pcDNA3.1-MAP3K1 (not including 3’-UTR) or (and) miR-451. The cell invasion was assessed using transwell assay; H: The expression of CDKN2D reversed G1 arrest of miR-451. Cells were transfected with pcDNA3.1-CDKN2D (not including 3’-UTR) or (and) miR-451. The cell cycle was assessed using flow cytometry assay; I: The expression of MAP3K1 did not reverse G1 arrest of miR-451. Cells were transfected with pcDNA3.1-MAP3K1 (not including 3’-UTR) or (and) miR-451. The cell cycle was assessed using flow cytometry assay (^aP < 0.05 vs control group).