Figure 4.

Regulation of EZH2 by hsa-miR-26a and hsa-let-7 family in CRC. (a) Schematic presentation showing miRNAs predicted to target EZH2 using TargetScan database. (b) EZH2 is predicted to be regulated by several members of the let-7 family, miR-26a-5p, and miR-363-3p, which were downregulated in colon cancer. (c) Schematic presentation depicting alignment of Let-7b-5p and miR-26a-5p mature sequence, and the putative binding sites within the 3′-UTR region of the EZH2 mRNA using TargetScan database. The exact positions of the interaction between EZH2 3′-UTR and both miRNA seed regions are indicated. Immunoblotting for EZH2 protein in HT115 transfected with the indicated pre-miRs at 48 h posttransfection. GAPDH was used as a loading control. (d) Inverse relationship between EZH2 and hsa-miR-26a-5p and hsa-let-7b-5p expression in 13 CRC and matched normal tissues. (e) siRNA-mediated knockdown of EZH2 (lower panel) or exogenous expression of hsa-let7-b-5p and hsa-miR-26a-5p (upper panel) led to significant reduction in cell viability in HT115 colon cancer cell. Data are presented as mean±S.E., n=12. *P<0.05; **P<0.005; ***P<0.0005