Fig. 5
Upregulation of miR-202-5p enhances the resistance of CML cells to Imatinib by inhibiting the apoptosis of CML cells. (a) K562 cells were transfected with miR-202-5p inhibitor or inhibitor-NC Cells and cultured in different times. CCK-8 assay was used to test the cell viability. *P < 0.05, **P < 0.01, **P < 0.001 vs. inhibitor-NC. (b) K562 cells were transfected with miR-202-5p inhibitor or inhibitor-NC for 48 h. Cell apoptosis were test by Annexin V-FITC/PI staining. **P < 0.01 vs. inhibitor-NC. (c) Stably express anti-miR-202-5p K562 cells or negative control K562 cells were injected subcutaneously in 200 μl 1640/Matrigel (100: 100) into the right posterior ankle of the nude mice to establish xenograft tumors (each group, n = 6). Tumor volumes were monitored by direct measurement with calipers and calculated by the formula: (length × width2)/2. ***P < 0.001 vs. LV-miR-NC. (d) qRT-PCR was used to detect miR-202-5p level in K562 cells, K562G cells and PBMCs of healthy donors. ** P < 0.01, *** P < 0.001 vs normal, ## P < 0.01 vs. K562 cell. (e) FISH assay was used to analyzed miR-202-5p level and localization in K562 cells and K562G cells compared with PBMCs of healthy donors. Scale bar = 20 μm. (f) qRT-PCR was used to detect miR-202-5p level in PBMCs of CML-CP (n = 6) patients and PBMCs of CML-BC patients (n = 6) *P < 0.05 vs. CML-CP. (g) K562G cells were transfected with miR-202-5p inhibitor or inhibitor-NC and then treated with Imatinib (3 μM) for 48 h. Cell apoptosis were evaluated by Annexin V-FITC/PI staining. **P < 0.01 vs. inhibitor NC. (h) K562G cells were transfected with miR-202-5p inhibitor or inhibitor-NC and then treated with Imatinib in different concentrations for 48 h. Imatinib IC50 of K562G cells was evaluated by CCK-8 assay. *P < 0.05 vs inhibitor-NC. (i) K562 cells were transfected with miR-202-5p mimic or mimic and then treated with Imatinib (0.1 μM) for 48 h. Cell apoptosis were evaluated by Annexin V-FITC/PI staining. *P < 0.01 vs. mimic-NC. (j) K562 cells were treated with Imatinib in different concentrations for 48 h. Imatinib IC50 of K562 cells was evaluated by CCK-8 assay. **P < 0.01 vs mimic-NC