微小RNA-328对人宫颈癌顺铂耐药HeLa细胞株耐药性的影响

Effect of microRNA-328 on the drug resistance of human cervical cancer cisplatin-resistant HeLa cell line

目的探讨微小RNA-328(miRNA-328)对顺铂(Cisplatin,DDP)耐药人宫颈癌HeLa细胞的逆转作用及相关的分子机制。方法体外用低浓度梯度递增方法构建顺铂耐药细胞株(HeLa/DDP);将细胞分为空白对照组(不做处理)、NC组(采用脂质体处理)、inhibitor组(转染NC序列)和mimics组(转染miR-328 mimics)。用细胞计数试剂盒-8(CCK-8)实验检测DDP处理后各组细胞增殖情况;用流式细胞术检测细胞凋亡情况;用Transwell小室检测细胞迁移情况;用蛋白质印迹法检测相关蛋白表达量。结果miRNA-328在HeLa/DDP细胞中处于低表达状态。空白对照组、NC组、inhibitor组及mimics组在顺铂作用下的的IC50分别为28.5,29.1,66.2,10.2μg·mL^(-1),耐药指数分别为2.5,2.6和5.8及0.9;迁移细胞比例分别为(32.5±3.6)%,(27.6±2.4)%,(49.6±3.1)%及(13.1±1.1)%;凋亡率分别为(4.87±0.89)%,(6.83±0.57)%,(1.11±0.18)%,(40.27±2.18)%,mimics组与空白对照组、NC组、inhibitor组比较,差异均有统计学意义(均P<0.05)。结论miRNA-328过表达能够降低HeLa/DDP对顺铂的耐药性,提高其对顺铂的敏感性。

Objective To explore the reversal effect of microRNA-328(miRNA-328)on cisplatin(DDP)-resistant human cervical cancer HeLa cells and related molecular mechanisms.Methods The cisplatin-resistant cell line(Hela/DDP)was constructed in vitro by low concentration gradient method;the cells were divided into blank control group(no treatment),NC group(treated with lipidosome),inhibitor group(transfected NC sequences)and mimics group(transfected miR-328 mimics).The cell counting kit-8(CCK-8)experiment was used to detect the cell proliferation of the three groups after DDP treatment.Flow cytometry was used to detect cell apoptosis.Transwell chamber was used to detect cell migration and invasion.Western blot was used to detect related protein expression.Results The IC_(50) of blank control group,NC group,inhibitor group,mimics group of cisplatin were 28.5,29.1,66.2 and 10.2μg·mL^(-1);the resistance index were 2.5,2.6,5.8 and 0.9;the proportions of migrating cells were(32.5±3.6)%,(27.6±2.4)%,(49.6±3.1)%and(13.1±1.1)%;apoptosis rates were(4.87±0.89)%,(6.83±0.57)%,(1.11±0.18)%,(40.27±2.18)%.There were statistically significant differences between mimics group and blank control group,NC group and inhibitor group(all P<0.05).Conclusion Overexpression of miRNA-328 can reduce the resistance of HeLa/DDP to cisplatin,increase its sensitivity to cisplatin.