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. 2016 Jan 30;7(8):8783–8796. doi: 10.18632/oncotarget.7088

Figure 5. miR-1827 enhances p53-mediated apoptosis and senescence in response to stress.

Figure 5

(A–C) miR-1827 enhances p53-mediated apoptosis. (A) miR-1827 reduced MDM2 protein levels and increased p53 protein levels in HCT116 cells treated with 5-FU. p53+/+ and p53−/− HCT116 cells transfected with miR-1827 mimic or miR-con were treated with 5-FU for 12 h for western-blot assays. (B) miR-1827 enhanced p53-mediated apoptosis in HCT116 and RKO cells treated with 5-FU. (C) miR-1827 inhibitor reduced p53-mediated apoptosis in HCT116 cells treated with 5-FU. In B & C, cells were transfected with miR-1827 mimic or miR-con (B), or transfected with miR-1827 inhibitor or control inhibitor (C), and were then treated with 5-FU (300 μM for HCT116 cells and 500 μM for RKO cells, respectively). Apoptosis was measured by Annexin V staining in a flow cytometer at 36 h after treatment. Data are presented as mean ± SD (n = 4). #p < 0.05; *p < 0.01; **p < 0.001; Student t-test. (D–G) miR-1827 enhances p53-mediated senescence. (D) miR-1827 reduced MDM2 protein levels and increased p53 protein levels in HCT116 cells treated with Doxorubicin (Dox). p53+/+ and p53−/− HCT116 cells transfected with miR-1827 mimic or miR-con were treated with Doxorubicin for 12 h for western-blot assays. (E) miR-1827 enhanced p53-mediated senescence in HCT116 cells treated with Doxorubicin. (F) miR-1827 enhanced p53-mediated senescence in RKO cells treated with Doxorubicin. (G) miR-1827 inhibitor reduced p53-mediated senescence in HCT116 cells treated with Doxorubicin. In E–G, cells transfected with miR-1827 mimic (E, F) or miR-1827 inhibitor (G) were treated with Doxorubicin (100 nM for HCT116 cells and 300 nM for RKO cells, respectively) for 3 days before cellular senescence was detected by SA-b-gal staining. Left panels in E: represented images of SA-β-gal staining of senescent cells. Scale bar, 100 μm. E (Right panels) and F, G: the percentage of SA-β-gal positive cells. Data are presented as mean ± SD (n = 3). #p < 0.05; **p < 0.001; Student t-test.