Figure 3

HHT regulated mature miR-370 level in K562 cells. K562 cells were incubated with HHT or PBS and harvested after 72 or 96 h. Real-time PCR analysis of the mRNA expression of mature miR-370 and FoxM1. Western blot analysis of FoxM1 protein level. (A) HHT increased the expression of mature miR-370 in a dose-dependent manner. The mRNA level of FoxM1 decreased in K562 cells with HHT incubation, **P < 0.01 vs control, Data are mean ± SEM of 3 independent experiments. (B) HHT regulated the expression of miR-370 and FoxM1 time-dependently. *P < 0.05, **P < 0.01 vs control, Data are mean ± SEM of 3 independent experiments. (C-D) Western blot analyis of protein level of FoxM1 after HHT treatment at different concentrations or different times. β-actin was a loading control. (E) The protein expression of FoxM1 with incubation of HHT and miR-370 inhibitor. The results were confirmed by 3 independent experiments. (F) Anti-miR-370 treatment partially reversed HHT-induced apoptosis. Cells were treated with 0.015 μM HHT + NC or 0.015 μM HHT + miR-370 inhibitor for 72 h. Flow cytometry of K562 cells by double-staining with PI and Annexin V. The proportion of apoptotic cells is shown in the upper right corners of Q2 and Q4 cells. The results are from 3 independent experiments. (G) Mature miR-370 expression was downregulated in K562 cells treated with HHT + miR-370 inhibitor, **P < 0.01 vs HHT + NC group. FoxM1 mRNA level was upregulated, *P < 0.05 vs HHT + NC group. Data are mean ± SEM of 3 independent experiments.