miR-100在上皮性卵巢癌顺铂耐药细胞中的表达及其与耐药的关系

Expression of microRNA-100 and its correlation with drug resistance in human ovarian cancer SKOV3/DDP cells

目的研究微小RNA-100(mi R-100)在人上皮性卵巢癌顺铂耐药细胞中的表达及其与顺铂耐药的关系。方法脂质体lipofectamine 2000介导成熟mi R-100模拟物(mimics)及阴性对照片段(NC)、阻遏物(inhibitor)及阴性对照片段(inhibitor NC)分别转染人卵巢癌顺铂耐药细胞株SKOV3/DDP。实验分为6组:SKOV3组、SKOV3/DDP组、mi R-100 mimics组、NC组、mi R-100 inhibitor组及inhibitor NC组。实时荧光定量PCR及CCK8法分别检测各组细胞中mi R-100的表达和顺铂半数抑制浓度(IC50)。结果(1)SKOV3/DDP细胞的顺铂耐药指数(RI)为2.23;(2)mi R-100在SKOV3/DDP细胞中低表达,与SKOV3相比差异有统计学意义(P<0.001);(3)转染mi R-100 mimics后,SKOV3/DDP细胞中mi R-100的表达水平与NC组相比上升38.29倍,差异有统计学意义(P<0.01);mi R-100 mimics组顺铂IC50与NC组相比明显降低,差异有统计学意义(P<0.001);(4)转染mi R-100 inhibitor后,SKOV3/DDP细胞中mi R-100的表达水平与inhibitor NC组相比下降97.7%(P<0.001);mi R-100 inhibitor组顺铂IC50与inhibitor NC组相比明显升高(P<0.001)。结论 mi R-100在人卵巢癌顺铂耐药细胞中低表达,mi R-100可增加卵巢癌顺铂耐药细胞株对顺铂的敏感性,从而逆转其耐药。

Objective To investigate the expression of micro RNA-100（mi R-100） and the relationship with cisplatin resistance in human ovarian epithelial cancer SKOV3/DDP cells. Methods The SKOV3/DDP cells were transfected with the mimics or inhibitor of mi R-100 or negative control RNA（NC） or inhibitor negative control RNA（inhibitor NC） by lipofectamine 2000.The experiment was divided into six groups： SKOV3 group, SKOV3/DDP group, mi R-100 mimices group, NC group, mi R-100 inhibitor group and inhibitor NC group. The expression of mi R-100 and the cisplatin IC50 were measured by real-time PCR and CCK8 assay respectively. Results（1）The cisplatin resistance index of SKOV3/DDP was 2.23;（2）The express level of mi R-100 in SKOV3/DDP cells was significantly lower than that in SKOV3 cells（P〈0.001）;（3）After transfected with mi R-100 mimics,SKOV3/DDP cells showed that the level of mi R-100 was 38.29 times higher than that in the NC group（P〈0.01）. The cisplatin IC50 of mi R-100 mimices group was significantly lower than that in the NC group（P〈0.001）;（4） After transfected with mi R-100 inhibitor, the level of mi R-100 0f SKOV3/DDP was decreased by 97.7%. The cisplatin IC50 of mi R-100 inhibitor group was significantly increased as compared with that in the inhibitor NC group（P〈0.001）. Conclusion The expression of mi R-100 is downregulated in SKOV3/DDP cells. Overexpressing mi R-100 may effectively increase the sensitivity to cisplatin of human ovarian epithelial cancer SKOV3/DDP cells and may reverse cisplatin-resistance of EOC（epithelial ovarian cancer）.