Fig. 2

CCNG1 is the direct target of miR-9 and modulates the paclitaxel sensitivity of EOC cells a. Modulating miR-9 expression changed paclitaxel sensitivity of ST30 and SKOV3 cells. ST30 cells were transfected with miR-9 mimic or negative control(IC50 = 820.89 ± 21.62 nM VS 2424.56 ± 56.83nM, P = 0.001), SKOV3 cells were transfected with miR-9 inhibitor or negative control (IC50 = 122.74 ± 10.12 nM VS64.63 ± 2.74 nM, P = 0.000), the cytotoxicity of paclitaxel on EOC cells were assessed by MTS assay. b. Western blot analysis of CCNG1 in ST30 cells transfected with miR-9 mimic or negative control. GAPDH was used as house-keeping gene. c. Dual luciferase reporter assay. 293 T cells were transfected with CCNG1 -wild type 3′UTR vectors or mutant 3′UTR vectors together with miR-9 mimic or its negative control. Luciferase activity was measured 48 h after cotransfection. A decrease of the luciferase activity was observed in miR-9 overexpressing cells compared with control (* P = 0.008). d. Modulating miR-9 expression changed paclitaxel sensitivity of A2780 and A2780R cells. A2780 cells were transfected with miR-9 inhibitor or negative control (IC50 = 95.644 ± 12.03 nM VS 38.16 ± 6.18 nM, P = 0.000), A2780R cells were transfected with miR-9 mimic or negative control(IC50 = 194.94 ± 9.36 nM VS 774.03 ± 49.19 nM, P = 0.002). e. Western blot analysis of CCNG1 in SKOV3 cells transfected with miR-9 inhibitor, negative control or inhibitor combined with CCNG1 siRNA. GAPDH was used as house-keeping gene. f. Modulating CCNG1 expression changed paclitaxel sensitivity of ovarian carcinoma. Knockdown of CCNG1 alone enhanced paclitaxel cytotoxcity to ST30 cells (IC50 = 1468.50 ± 32.19 nM VS 2545.84 ± 168.83 nM, P = 0.000), while deleption CCNG1 reversed the role of miR-9 inhibitor on the paclitaxel sensitivity of SKOV3 cells (IC50 = 65.35 ± 13.47 nM VS 177.36 ± 20.88 nM, P = 0.001). The experiments were repeated three times