miR-21 Targets ASPP2 to Inhibit Apoptosis via CHOP-Mediated Signaling in Helicobacter pylori-Infected Gastric Cancer Cells

Figure 4

H. pylori induces ASPP2 downregulation and inhibition of apoptosis in HPGC. (a–c) AGS and MKN45 cells transfected with mock and ASPP2-overexpressing vectors were incubated with H. pylori at 30 and 90 MOI or PBS for 32 h. (a) ASPP2 protein levels in the cell lysates were measured via immunoblotting. (b) The percentage of apoptotic cells was analyzed by TUNEL assay. (c) The percentage of early and late apoptotic cells was assessed via Annexin V/PI assay. All the data were presented as mean ± SD of 3 replicates. (d) Relative cleaved caspase 3 expression levels in HPGC tumor tissues (n = 35) as compared to adjacent normal tissues (n = 35) by IHC and the correlation with ASPP2 levels. Upper left, representative images were captured at 200x magnification. Scale bar, 50 μm. Lower left, relative quantitation of cleaved caspase 3 levels by H score ranking. The significance of differences in H score between two groups was compared using the nonparametric Mann–Whitney U test. Right, Spearman correlation analysis between ASPP2 and cleaved caspase 3 levels in HPGC tumor tissues (n = 35).  < 0.05,  < 0.01, and  < 0.001.

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