微小RNA-98-5p靶向调控核糖核苷酸还原酶小亚基M2调控宫颈癌细胞顺铂的耐药性

目的 探讨微小RNA（miR)-98-5p对顺铂(DDP)耐药宫颈癌细胞顺铂敏感性的调控及其机制。 方法 用脂质体法将DDP+miR-NC组（转染miR-NC）、DDP+miR-98-5p组（转染miR-98-5p mimics）、DDP+si-NC组（转染si-NC）、DDP+si-核糖核苷酸还原酶小亚基M2（RRM2）组（转染si-RRM2）、DDP+miR-98-5p+pcDNA组（共转染miR-98-5p mimics和pcDNA）、DDP+miR-98-5p+pcDNA-RRM2组（共转染miR-98-5p mimics和pcDNA-RRM2）转染至HeLa/DDP组细胞；Real-time PCR、Western blotting、CCK-8、迁移实验（Transwell）和双荧光素酶报告基因检测细胞中miR-98-5p、RRM2、细胞周期蛋白D1（cyclin D1）、P21、基因金属蛋白酶（MMP）-2、MMP-9的表达、细胞的抑制率、半数抑制浓度（IC50）、迁移和侵袭及荧光活性。 结果 与HeLa组细胞相比，HeLa/DDP组细胞中miR-98-5p表达显著降低，RRM2表达显著升高，IC₅₀值显著升高（P<0.05）；过表达miR-98-5p或抑制RRM2可明显抑制HeLa/DDP细胞的增殖、迁移和侵袭，下调cyclin D1、MMP-2、MMP-9蛋白，上调P21蛋白；miR-98-5p明显抑制野生型RRM2细胞的荧光活性，并且过表达RRM2反转了miR-98-5p对HeLa/DDP细胞增殖、迁移侵袭的抑制作用。 结论 MiR-98-5p可抑制顺铂耐药宫颈癌细胞的增殖、迁移和侵袭，增强对顺铂的敏感性，其机制与靶向RRM2相关，将可为顺铂耐药宫颈癌细胞的治疗提供方向。

MicroRNA-98-5p targeting ribonucleotide reductase small subunit M2 regulating cisplatin resistance in cervical cancer cells

Objective To investigate the regulation and mechanism of microRNA(miR)-98-5p on cisplatin sensitivity in cisplatin resistant cervical cancer cells. Methods The cisplatin(DDP)+miR-NC group (transfected miR-NC), DDP+miR-98-5p group (transfected miR-98-5p mimics), DDP+si-NC group (transfected si-NC), DDP+si-ribonucleotide reductase subunit M2(RRM2) group (transfected si-RRM2), DDP+miR 98-5p+pcDNA group (co-transfected miR-98-5p mimics and pcDNA), DDP+miR-98-5p+pcDNA-RRM2 group (co-transfected miR-98-5p mimics and pcDNA-RRM2) were transfected into HeLa/DDP cells by liposome method . Real-tim PCR, Western blotting, CCK-8, Transwell chamber and dual luciferase reports gene detection assay were used to detect the expression of miR-98-5p, RRM2, cyclin D1, P21, matrix metalloproteinase(MMP)-2 and MMP-9 in cells, inhibition rate, half inhibitory concentration(IC₅₀), migration and invasion and fluorescence activity. Results Compared with the HeLa group, the expression of miR-98-5p was significantly decreased in HeLa/DDP group, the expression of RRM2 was significantly increased, the IC50 value was significantly increased (P<0.05). Overexpression miR-98-5p or inhibition RRM2 could significantly inhibit the proliferation, migration and invasion of HeLa/DDP cells, down-regulated proteins expression of cyclin D1, MMP-2 and MMP-9, and up-regulated proteins expression of P21; miR-98-5p inhibited the fluorescence activity of wild-type RRM2 cells, and overexpress RRM2 could reverse the inhibitory effect of miR-98-5p on proliferation, migration and invasion of HeLa/DDP cells. Conclusion miR-98-5p can inhibit the proliferation, migration and invasion of cisplatin-resistant cervical cancer cells and enhance the sensitivity to cisplatin. The mechanism is related to the targeting of RRM2, which will provide directions for the treatment of cisplatin-resistant cervical cancer cells.