Figure 5.
Metadherin (MTDH) was a downstream target gene of miR‐125a. (a) Prediction of binding sites between miR‐125a and 3′ UTR of MTDH (in the box) were shown according to TargetScan tools. The corresponding mutant of MTDH 3′ UTR (MUT‐MTDH) was also presented. (b) Luciferase activity of wild‐type of MTDH (WT‐MTDH) and MUT‐MTDH was confirmed by dual luciferase reporter assay in HCC‐70 and MB231 cells when cotransfected with miR‐125a or miR‐control. HCC‐70 (
) miR‐control and (
) miR‐125a. MB231 (
) miR‐control and (
) miR‐125a. (c) RT‐qPCR measured MTDH mRNA level in breast cancer tissues (n = 45) compared with the paired normal tissues. (d) Spearman's rank correlation analysis clarified the association between miR‐125a and MTDH expression in breast cancer tissues (n = 45). (e, f) RT‐qPCR and western blotting measured MTDH levels in breast cancer cell lines (HCC‐70 and MB231) comparing to MCF‐10A. (g) RT‐qPCR determined the transfection efficiency of miR‐125a inhibitor (anti‐miR‐125a) and its control (anticontrol) in HCC‐70 and MB231 cells. (
) anticontrol and (
) anti‐miR‐125a. (h, i) RT‐qPCR and western blotting detected MTDH expression levels in HCC‐70 and MB231 cells when transfected with miR‐125a, miR‐control, anti‐miR‐125a and anticontrol. (
) miR‐control, (
) miR‐125a, (
) anticontrol and (
) anti‐miR‐125a. Data represent mean ± SEM and *P < 0.05.