Fig. 3

Twist is a direct target of miR-548c in EC and OC cells. a In silico prediction and molecular pathway enrichment analysis indicate that miR-548c-regulated genes are preferentially involved in pathway associated with tumor growth and metastasis. The top-20 ranking KEGG pathways were listed. b The putative conserved miR-548c-binding site in the Twist 3’-UTR. Mutation was generated in the binding site. c Western blotting analysis of Twist expression in RL95-2, HEC-1 or SKOV-3 cells after knockdown or overexpression of miR-548c. d Indicated cells were transfected with reporter constructs containing either wild-type (WT) Twist, or Twist 3’-UTR with mutation (Mut), along with miR-548c mimic, anti-miR-548c inhibitor or negative controls, respectively. Relative luciferase activity was assayed. e qPCR analysis of EMT, invasion and stemness-related genes in RL95-2 upon miR-548c knockdown, or in HEC-1 or SKOV-3 cells after overexpression of miR-548c. ^** P < 0.01