Figure 4.

CircHGF inhibited the proliferation and osteogenic differentiation of BMSCs in ONFH

(A) The BMSCs were treated with si-circRNA or its control si-NC, and the cell proliferation ability was revealed by the cell-cycle analysis, with the cell population of G1, G2, and S cell-cycle phase indicated. ^#p < 0.05 versus si-NC group for comparing cell percentage in S phase, ^&p < 0.05 versus si-NC group for comparing cell percentage in G1 phase, n = 3. (B) The BMSCs were treated with circHGF over-expression vector OE-circRNA or its control OE-NC, and the cell proliferation ability was revealed by the cell-cycle analysis. ^#p < 0.05 versus OE-NC group for comparing cell percentage in S phase, ^&p < 0.05 versus OE-NC group for comparing cell percentage in G1 phase, n = 3. (C and D) The protein expression levels of osteogenic marker genes in BMSCs were detected by western blot analysis. GAPDH served as an internal control correspondingly. The quantitative analysis showed the protein expression levels of ALP, BMP2, RUNX2, and OCN were all increased in the si-circRNA group compared with the si-NC group and were decreased in the OE-circRNA group compared with the OE-NC group. (E) The osteogenic differentiation ability of BMSCs at the 7th and 14th day after si-circRNA or control treatment was determined by alizarin red staining. (F) The osteogenic differentiation ability of BMSCs at the 7th and 14th day after OE-circRNA or OE-NC treatment was evaluated by alizarin red staining. Data were shown as mean ± SD. ^∗∗p < 0.01, n = 3. Data among multiple groups were analyzed by one-way ANOVA test.