miR-181a在卵巢癌细胞中对顺铂的耐药作用

Studies on Cisplatin Resistance of miR-181a in Ovarian Cancer Cells

目的探讨miR-181a在卵巢癌化疗抵抗方面的影响及其机制。方法qRT-PCR检测卵巢癌细胞A2780及A2780/DDP中miR-181a的表达;对A2780及A2780/DDP细胞分别进行miR-181a mimics和inhibitors的转染,并利用qRT-PCR技术验证;MTT法检测转染前后细胞对顺铂的敏感性,利用TargetScan、miRDB与miRwalk 3个MicroRNA靶基因数据库预测miR-181a下游靶点,并通过Western blot技术分析预测基因的表达。结果与A2780细胞相比,miR-181a在A2780/DDP细胞中的表达明显减弱(P<0.05)。干扰miR-181a表达后,A2780细胞对顺铂的抵抗作用减弱(P<0.05),而上调miR-181a表达,A2780/DDP细胞对顺铂的抵抗作用增强(P<0.05)。通过TargetScan、miRDB、miRwalk三个数据库预测到PRKCD可作为miR-181a下游靶基因,下调miR-181a表达可使PRKCD蛋白表达明显增强(P<0.05);反之,上调miR-181a表达能够显著抑制PRKCD蛋白表达(P<0.05)。结论miR-181a卵巢癌细胞对顺铂的耐药性方面具有抑制作用。

Objective To investigate the effect of miR-181 a on chemotherapy resistance of ovarian cancer and its regulatory mechanism.Methods Real-time PCR was used to detect the expression of miR-181 a in A2780 and cisplatin resistant cell lines(A2780/DDP),and siltation/overexpression changed the expression of miR-181 a in A2780 and A2780/DDP cells.MTT method was used to determine the sensitivity of cells to cisplatin before and after the transfection.Three MicroRNA target gene databases,TargetScan,miRDB and miRwalk,were used to predict the downstream targets of miR-181 a,and the changes in protein expression of target genes were analyzed by Western blot.Results Compared with A2780 cells,the expression of miR-181 a in A2780/DDP cells was significantly decreased(P<0.05).After the transfection of A2780 cells with miR-181 a inhibitor(interfering with Mir-181 A expression),the sensitivity of A2780 cells to cisplatin decreased(P<0.05),and after the transfection of A2780/DDP cells with miR-181 a mimic(overexpressing miR-181 a),The sensitivity of cells to cisplatin was increased(P<0.05).It was predicted that PRKCD could be used as the downstream target gene of miR-181 a by TargetScan,miRDB and miRwalk databases,and down-regulation of miR-181 a expression could significantly enhance PRKCD protein expression(P<0.05).On the contrary,up-regulation of miR-181 a significantly inhibited PRKCD protein expression(P<0.05).Conclusion mi R-181 a may inhibit cisplatin resistance of ovarian cancer cell A2780 by regulating PRKCD.