Research Paper Volume 14, Issue 7 pp 3105—3128

Figure 3. Circ_6014 regulated the expression of miR-885-3p. (A) MiRNAs that have binding sites with circ_6014 according to two databases (miRanda and RNA hybrid). (B, C) The expression level of miR-885-3p increased (B) or decreased (C) with the down- or upregulation of circ_6014 expression. (D) The predicted wild-type binding sites and designed mutant-type binding sites of miR-885-3p and circ_6014. (E, F) Dual-luciferase reporter assays conducted in HEK293T and MDA-MB-231 cells with miR-885-3p and circ_6014 suggested that miR-885-3p decreased the relative activity of luciferase reporters of the wild-type group. (G, H) Transfection of si-NC + miRNA-inhibitor-NC, si- circ_6014 + inhibitor-miR-885-3p or OE-NC + miRNA-mimics-NC, or OE- circ_6014 + miR-885-3p and the absorbance of formazan solution at 490 nm showed that the decrease or increase in miR-885-3p could restrain the impact caused by down- or upregulating circ_6014 expression. (I, J) Colony formation of both cell lines in six-well plates transfected with si-NC + miRNA-inhibitor-NC, si-circ_6014 + inhibitor-miR-885-3p or OE-NC + miRNA-mimics-NC, or OE-circ_6014 + miR-885-3p. (K, L) Columns used to compare the difference between cell counting numbers per area revealed that miR-885-3p could reverse the changes in biological function caused by circ_6014. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.