Figure 4
From: MicroRNA-26a inhibits osteosarcoma cell proliferation by targeting IGF-1
IGF-1 is a direct target of miR-26a. (a) IGF-1 3’-UTR contains one predicted miR-26a binding site. The mutagenesis nucleotides are indicated in red. (b) Dual luciferase reporter assay. HEK293 cells were transfected with wild-type 3’-UTR-reporter or mutant (Mut) constructs together with miR-26a mimics or NCs. Relative firefly luciferase expression was normalized to Renilla luciferase. (c and d) qRT-PCR and Western blot to measure IGF-1 mRNA and protein level in MG-63 cells transfected with NC, miR-26a mimics, or inhibitors. (e and f) IGF-1 rescues the suppressive roles of miR-26a in GC cell proliferation. MG-63 cells expressing miR-26a mimics or NC were transfect with or without IGF1 plasmids. Cell proliferation analysis was performed by cell counting (e) and MTS assay (f). (g) The expression of IGF-1 in 32 pairs of osteosarcoma samples and matched adjacent noncancerous tissues by qRT-PCR. Data are presented as the lg2 fold-change in osteosarcoma tissues relative to the adjacent normal tissues. (h) The correlation between miR-26a expression and IGF-1 expression in osteosarcoma tissues. Data are presented as mean ± s.d. from at least three independent experiments. *P < 0.05; **P < 0.01.
