Figure 2
Anti-proliferative effects of miR-125b in MM cells. (a) qRT-PCR analysis of miR-125b-5p expression in SK-MM-1 cells transduced with either Lenti-miR-125b-1 or Lenti-miR-125b-2; the results are shown as average miR-125b-5p expression levels after normalization with RNU44 and ΔΔCt calculations. (b) CCK-8 proliferation assay of NCI-H929, SK-MM-1 and RPMI-8226 cells transduced with a lentivirus carrying either the miR-125b-1 (Lenti-miR-125b-1) or the miR-125b-2 (Lenti-miR-125b-2) genes; the effects on cell proliferation were assessed at 2, 3 and 4 days after selection by puromycin. qRT-PCR analysis of miR-125b-5p expression in SK-MM-1 cells transfected with either (c) miR-125b-5p inhibitors or (e) miR-125b-5p mimics; the results are shown as average miR-125b-5p expression levels after normalization with RNU44 and ΔΔCt calculations. (d) CCK-8 proliferation assay was performed 2–4 days after transfection of eight MM cell lines (MM.1S, MM.1R, U266/LR7, RPMI-8226, RPMI-8226/DOX40, SK-MM-1, KMS-12-BM and INA-6) with miR-125b-5p inhibitors or scrambled controls (miR-NC inhibitors). (f) CCK-8 proliferation assay was performed 2–4 days after transfection of 10 MM cell lines (NCI-H929, U266, MM.1S, MM.1R, U266/LR7, RPMI-8226, RPMI-8226/DOX40, SK-MM-1, KMS-12-BM and INA-6) with miR-125b-5p mimics or scrambled controls (miR-NC mimics). (g) CCK-8 assay of CD138+ cells from three different MM patients transfected with miR-125b-5p or miR-NC. The assay was performed 48 h after cell transfection. (h) 7-AAD flow cytometry assay was performed 48 h after transfection of PBMCs from six healthy donors (HDs) with miR-125b-5p or miR-NC. Data represent the average ±s.d. of three independent experiments. P-values were obtained using two-tailed t-test.
