Figure 2. CircRNA Cdr1as directly interacts with miR-1270. (A) Prediction results on RegRNA 2.0 and CircInteractome. (B) Expressions of miR-1270 and miR-671-5p in HCC cells and HL-7702 cells. MiR-1270 level was significantly reduced in HCC cells compared to HL-7702 cells. (C) qRT-PCR detection of the relative expression of miR-1270 in paired HCC tumor and paired para-carcinoma tissues (n=42). (D) Correlation between circRNA Cdr1as and miR-1270 in HCC samples. (E) Expressions of circRNA Cdr1as and miR-1270 in the nucleus and cytoplasmic fractions of HepG2 and SMMC-7721 cells were analyzed by qRT-PCR. (F) RIP experiment confirmed the binding relationships of circRNA Cdr1as and miR-1270 in HepG2 and SMMC-7721 cells. CircRNA Cdr1as and miR-1270 levels were detected using qRT-PCR. (G) Bioinformatics evidence for the binding of miR-1270 to the 3'-UTR of circRNA Cdr1as. (H) The luciferase activity in HepG2 and SMMC-7721 cells after co-transfection of plasmid (pGL3-circRNA Cdr1as-WT or pGL3-circRNA Cdr1as-MUT) and miRNA-1270 mimics tested by dual-luciferase reporter gene assay. (I) MiR-1270 co-localized with circRNA Cdr1as in HCC tissues or normal liver tissues was detected by FISH. Results were presented as mean ± SD. *P<0.05, **P<0.01, ***P<0.001. All of the experiments were performed in triplicate.