微小RNA-101-3p对卵巢癌细胞紫杉醇敏感性的影响

Mechanism of microRNA-101-3p enhancing paclitaxel sensitivity of ovarian cancer cells

目的探讨微小RNA-101-3p(miR-101-3p)调控Notch同源物1(Notch1)对卵巢癌细胞紫杉醇敏感性的影响。方法将A2780/Taxol细胞分为紫杉醇组(用1.8μmol·L^(-1)紫杉醇处理),第1转染组(转染miR-101-3p mimics,用1.8μmol·L^(-1)紫杉醇处理),第2转染组(转染pcDNA3.1-Notch1,用1.8μmol·L^(-1)紫杉醇处理),第3转染组(同时转染pcDNA3.1-Notch1和miR-101-3p mimics,用1.8μmol·L^(-1)紫杉醇处理)。用逆转录实时荧光定量聚合酶链反应(RT-qPCR)检测miR-101-3p在A2780和A2780/Taxol细胞中的表达水平。48 h后,用CCK-8检测细胞增殖活性。用流式细胞术检测细胞周期,用蛋白质印迹(Western blot)法检测Notch1的表达水平。结果miR-101-3p在A2780和A2780/Taxol细胞中的表达水平分别为1.00±0.04,0.35±0.05,差异有统计学意义(P<0.05)。紫杉醇组、第1转染组、第2转染组、第3转染组的细胞48 h增殖率分别为(141.82±5.45)%,(106.06±7.35)%,(184.24±8.20)%,(146.06±7.57)%;G0/G1期细胞百分比分别为(40.17±2.93)%,(53.38±2.56)%,(19.97±3.22)%,(35.49±4.51)%;S期细胞百分比分别为(36.74±4.07)%,(33.05±3.13)%,(40.12±3.29)%,(37.05±2.39)%;G2/M期细胞百分比分别为(23.09±1.42)%,(13.57±0.58)%,(39.92±2.68)%,(27.46±4.59)%;Notch1的相对表达水平分别为1.03±0.09,0.53±0.04,1.89±0.21,1.26±0.11。第1转染组、第2转染组分别与紫杉醇组比较,第3转染组与第2转染组比较,差异均有统计学意义(均P<0.05)。结论miR-101-3p在紫杉醇耐药细胞A2780/Taxol中低表达,过表达miR-101-3p可通过靶向下调Notch1的表达增强A2780/Taxol细胞紫杉醇敏感性。

Objective To investigate the effect of microRNA-101-3p(miR-101-3p)regulating Notch homolog 1(Notch1)on the sensitivity of ovarian cancer cells to paclitaxel.Methods The experiment was divided into paclitaxel group(treated with 1.8μmol·L^(-1) paclitaxel),transfection-1 group(transfected with miR-101-3p mimics and treated with 1.8μmol·L^(-1) paclitaxel),transfection-2 group(transfected with pcDNA3.1-Notch1 and treated with 1.8μmol·L^(-1) paclitaxel),and transfection-3 group(transfected with both pcDNA3.1-Notch1 and miR-101-3p mimics and treated with 1.8μmol·L^(-1) paclitaxel).Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-101-3p in A2780 and A2780/Taxol cells.Cell proliferative activity was detected by CCK-8.Cell cycle was detected by flow cytometry.Western blot was used to detect the expression level of Notch1.Results The expression of miR-101-3p in A2780 and A2780/Taxol cells were 1.00±0.04,0.35±0.05,respectively,with statistically significant difference(P<0.05).The cell proliferation rates of paclitaxel group,transfection-1 group,transfection-2 group,and transfection-3 group at 48 h were(141.82±5.45)%,(106.06±7.35)%,(184.24±8.20)%,(146.06±7.57)%,respectively.The percentage of cells at G0/G1 stage were(40.17±2.93)%,(53.38±2.56)%,(19.97±3.22)%,(35.49±4.51)%;S stage were(36.74±4.07)%,(33.05±3.13)%,(40.12±3.29)%,(37.05±2.39)%,G2/M stage were(23.09±1.42)%,(13.57±0.58)%,(39.92±2.68)%,(27.46±4.59)%,respectively.The relative expression levels of Notch1 were 1.03±0.09,0.53±0.04,1.89±0.21,and 1.26±0.11,respectively.There were significant differences between transfection-1 group and paclitaxel group,or transfection-2 group and paclitaxel group,or transfection-3 group and transfection-2 group(all P<0.05).Conclusion miR-101-3p was low expressed in paclitaxel resistant A2780/taxol cells.Overexpression of miR-101-3p could enhance paclitaxel sensitivity of A2780/taxol cells by down regulating Notch1 expression.