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 7. YY1 and eIF4A3 promoted circ-ZNF609 expression via linking to the circ-ZNF609 promoter and pre-mRNA, respectively. (A) YY1 sequence and binding sites (E1, E2 and E3) to circ-ZNF609 promoter region were predicted by using JASPAR database. (B) The efficiency detection of YY1 overexpression and siRNA transfection confirmed by qRT-PCR. (C) The circ-ZNF609 expression was detected in CCA with YY1 up-regulation and down-regulation by qRT-PCR. (D) ChIP assay was conducted to affirm the direct binding of YY1 to circ-ZNF609 promoter in QBC939 and CCLP-1 cells. (E) Schematic diagram of plasmids construction. (F) The luciferase activity of E3 wild type was markedly promoted by oe-YY1 co-transfection compared with controls in CCA cells. (G) The prediction results of circinteractome database indicated that eIF4A3 had binding sites in the upstream and downstream regions of circ-ZNF609 pre-mRNA. (H) The results of RIP demonstrated that eIF4A3 could directly bind to the circ-ZNF609 pre-mRNA in CCA cells. (I) Knockdown efficiency and amplification efficiency of eIF4A3 confirmed by qRT-PCR. (J) The expression of circ-ZNF609 in CCLP-1 and QBC939 transfected with high expression eIF4A3 and low expression eIF4A3 was detected by qRT-PCR. (K) Schematic diagram of circ-ZNF609 tumorigenesis mechanisms in cholangiocarcinoma. **P< 0.01; ***P< 0.001.