Figure 5 | Cell Death & Disease

Figure 5

From: Long noncoding RNA UCA1 induced by SP1 promotes cell proliferation via recruiting EZH2 and activating AKT pathway in gastric cancer

Figure 5

EZH2 and AKT pathway is involved in regulation of UCA1 on cyclin D1 expression. (a) EZH2 and AKT/GSK-3B/ cyclin D1 axis-related proteins were detected by immunoblotting, using extracts from the NCI-N87 transfected with pcDNA-UCA1 and SGC-7901 with UCA1 siRNAs and negative control cell, respectively. GAPDH is used as loading control. Overexpression of UCA1 in NCI-N87 cells significantly increased EZH2 expression and activated the AKT/GSK-3B/cyclin D1 axis. Downregulating UCA1 in SGC-7901 cells inbibited the activations of these proteins. (b) EZH2-specific inhibitor EZP005687 and P-AKT inhibitor LY294002 were used to culture NCI-N87 cells transfected with pcDNA-UCA1 and empty vectors. Effect of EZP005687 on NCI-N87 with pcDNA-UCA1 reflected in reduction of EZH2 and inactivation of AKT/GSK-3B/cyclin D1 and LY294002 showed the same effect on NCI-N87 cells with pcDNA-UCA1 compared with NCI-N87 cells with empty vectors. (c) CHIP assay was performed to determine the interaction between EHZ2 and cyclin D1 promoter. Anti-EZH2 significantly harbored more promoter fragments of cyclin D1 than anti-IgG both in NCI-N87 and SGC-7901 cells (**P<0.01). Error bars represent the mean±S.D. of triplicate experiments

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