Figure 5.

MiR-25-3p enhanced the proliferation and osteogenic differentiation ability of BMSCs

(A) The relative expression levels of miR-25-3p in ONFH-BMSCs and control BMSCs were examined by RT-qPCR analysis. (B) The BMSCs were transfected with miR-25-3p mimics or its control mimics NC, and the relative expression levels of miR-25-3p were determined by RT-qPCR analysis. (C) The BMSCs were treated with miR-25-3p inhibitor or its control inhibitor NC, and the relative expression levels of miR-25-3p were determined by RT-qPCR analysis. (D and E) The cell proliferation ability of BMSCs with corresponding treatment was revealed by the cell-cycle analysis, with the cell population of G1, G2, and S cell-cycle phase indicated. ^#p < 0.05 versus the corresponding NC group for comparing cell percentage in S phase; ^&p < 0.05 versus the corresponding NC group for comparing cell percentage in G1 phase; n = 3. (F and G) The protein expression levels of osteogenic marker genes in BMSCs of different groups were detected by western blot analysis. The quantitative analysis showed the protein expression levels of ALP, BMP2, RUNX2, and OCN were increased in the miRNA mimics group compared with the mimics NC group and were decreased in the miRNA inhibitor group compared with the inhibitor NC group. (H and I) The osteogenic differentiation ability of BMSCs at the 7th and 14th day after corresponding treatment was evaluated by alizarin red staining. Data were shown as mean ± SD. ^∗∗p < 0.01, ^∗∗∗p < 0.001, n = 3. Data between two groups were analyzed by Student's t test. Data among multiple groups were analyzed by one-way ANOVA test.