Figure 4.

ZNRF2 functions as a direct target of hsa-miR-105 in epithelial ovarian cancer (EOC) cells. (A) Prediction of putative target genes of hsa-miR-105 by three online programs. (B) Transcriptional expression levels of 17 candidate genes in four pair of paclitaxel (PTX)-responsive and PTX-resistant EOC tissues were analyzed using real-time RT-qPCR (*P<0.05). (C and D) EOC cells were transfected with hsa-miR-105 mimics or Ctrl for 48 h, followed by RT-qPCR (*P<0.05 and **P<0.01 when compared to NC) or western blot analysis. (E) Representative immunostaining of ZNRF2 and hsa-miR-105 ISH in PTX-responsive and PTX-resistant EOC tissues. Scale bar, 25 µm. (F) Luciferase reporter assay with co-transfection of wild-type (WT) or mutant (Mu) ZNRF2 and hsa-miR-105 mimics in 293T cells (**P<0.01 when compared to Ctrl). (G) Ectopic overexpression of Flag-Ago2 with indicated miRNAs in 293T cells. Flag antibody was used to precipitate miRNA-Ago2-mRNA complex. The expression of ZNRF2 mRNA in complex with hsa-miR-105 was determined by PCR. ZNRF2, zinc and ring finger 2.