Figure 5. miR-125a-5p suppresses tumorigenesis through inhibition of HDAC4.

R2N1d breast cancer cell line was transfected with 5 μg of each indicated plasmid or siRNAs, or anti-miR-125a-5p (150 nmol/L). Cell growth was evaluated by XTT assay (A, B). Invasiveness was evaluated in a transwell invasion chamber (C, D), and migration was evaluated with a wound-healing assay (E, F). Data as the means ± SD were based on three independent experiments. *,P < 0.05 vs. untreated control as determined by two-tailed Student's t test. Scale bar = 200 um. (G, H) R2N1d-YFP cells were injected into the right sides of immunodeficient SCID mice (n = 6 per group). On day 7 after transplantation, mice were then treated with the indicated plasmids and reagents via intratumoral injection (10mg/kg; once a week injection). Tumor development was assessed by whole body bioluminescence imaging (top) of the 28 days after transplantation, and the mean fluorescence intensity was quantitated using the MetaMorph software (bottom). Scale bar = 2.5 cm. (I) R2N1d-YFP cells were mixed with matrigel and injected into the left lateral thorax of mice as in G and H. The extra- and intra-thoracic lymph nodes in the right lung were quantified with a dissecting microscope and pathologically confirmed by H&E staining.