Research Paper Volume 13, Issue 23 pp 25195—25212

YY1 and eIF4A3 are mediators of the cell proliferation, migration and invasion in cholangiocarcinoma promoted by circ-ZNF609 by targeting miR-432-5p to regulate LRRC1


Figure 4. Circ-ZNF609 was involved in CCA progression via sponging miR-432-5p. (A) Detection of relative expression of circ-ZNF609 in cytoplasm and nucleus of CCA cells by qRT-PCR. (B) Targetscan, circinteractome and mirMap were used to identify potential miRNA target genes for circ-ZNF609. (C) Relative expression of potential target genes in CCA cells transfected with si-circ-1 and si-circ-2. (D) qRT-PCR was used to detect the relative expression of c miR-432-5p in CCA tumor tissues and adjacent normal tissues. (E) qRT-PCR was used to detect the correlation between the expression of circ-ZNF609 and miR-432-5p in CCA specimens. (F) The relative expression of miR-432-5p in CCA cells detected by qRT-PCR. (G) The efficiency detection of miR-432-5p mimics and inhibitor transfection confirmed by qRT-PCR. (H) Diagrammatic sketch of the binding sites between miR-432-5p and circ-ZNF609. (I) Luciferase activity in CCA cells after transfection of circ-ZNF609 WT or circ-ZNF609 MUT, miR-432-5p mimics or NC mimics. (J) RIP assay was utilized to further demonstrate the direct binding between miR-432-5p and circ-ZNF609. (K) Colony formation assay was used to detect the proliferation ability after miR-432-5p down-regulation and up-regulation. (L, M) The migration and invasion changes of tumor cells after miR-432-5p down-regulation and up-regulation detected by transwell. **P< 0.01; ***P< 0.001.