miR-17-5p Downregulation Contributes to Paclitaxel Resistance of Lung Cancer Cells through Altering Beclin1 Expression
Figure 4
BECN1 is a direct target of miR-17-5p.
(A) Schematic representation of the 3′- UTR of human BECN1 transcript. Predicted miR-17-5p binding site was depicted. The numbers (+135–141) represented the nucleotides that were predicted to base pair with the miR-17-5p seed sequence. (B) miR-17-5p directly binds to the 3′ UTR of BECN1 gene in A549-T24 cells. A549-T24 cells were co- transfected with renilla luciferase reporter plasmids (pCI-neo-RL-Bec-3′UTR-wt or pCI-neo-RL-Bec-3′UTR-mut), firefly luciferase plasmids (pGL3-FF) and pre-miR-17-5p precursor or anti-miR-17-5p or pre-miR-negative control precursor RNA using lipofectamine2000 transfection reagent. After 48 h, cells were harvested and lysed with passive lysis buffer. Luciferase activity was measured by using Promega dual luciferase reporter assay kit. The results were represented as relative fold repression (Renilla luc/Firefly luc activity) compared to control cells. (*p<0.05, **p<0.03 vs control, n = 4).