MicroRNA-31-3p靶向调控Sema4C表达对宫颈癌顺铂耐药性的作用及其机制研究

Effect of microRNA-31-3p targeted regulation of Sema4C expression on cisplatin resistance in cervical cancer and its mechanism

摘要:

目的 探讨人宫颈癌细胞microRNA-31-3p（miR-31-3p）的表达及其靶向调控Sema4C表达对顺铂耐药性的作用机制研究。方法 取对数生长期Hela细胞，分别转染miR-31-3p mimics（miR-31-3p mimics组）、空白质粒（对照组），以及在转染miR-31-3p mimics的基础上过表达Sema4C（miR-31-3p mimics＋Sema4C组）。采用实时荧光定量聚合酶链反应（qRT-PCR）检测miR-31-3p的表达，Western blotting检测转染mimics后Hela细胞Sema4C蛋白的表达，CCK-8法检测不同浓度顺铂作用下Hela细胞的耐药性变化。结果 miR-31-3p在宫颈癌细胞Hela中低表达（P <0.05）。miR-31-3p mimics组、miR-31-3p mimics+Sema4C组miR-31-3p相对表达量高于对照组（P <0.05），且miR-31-3p mimics组与miR-31-3p mimics+Sema4C组miR-31-3p相对表达量比较，差异无统计学意义（P >0.05）。miR-31-3p mimics组、miR-31-3p mimics+Sema4C组Sema4C蛋白相对表达量低于对照组（P <0.05）；miR-31-3p mimics+Sema4C组Sema4C蛋白相对表达量高于miR-31-3p mimics组（P <0.05）。不同浓度顺铂作用下，miR-31-3p mimics组Hela细胞生存率均低于对照组（P <0.05）；2 μmol/L、3 μmol/L、4 μmol/L、5 μmol/L顺铂作用下，miR-31-3p mimics+Sema4C组Hela细胞生存率高于miR-31-3p mimics组（P <0.05）。结论 miR-31-3p靶向调控Sema4C表达，降低宫颈癌细胞对顺铂的耐药性。

Abstract:

Objective To investigate the expression of microRNA-31-3p (miR-31-3p) in human cervical cancer cells and miR-31-3p's regulation of transmembrane signaling protein 4C (Sema4C) expression to reduce the resistance of cervical cancer cells to cisplatin.Methods MiR-31-3p mimics were transfected into Hela cells, and the expression of miR-31-3p was detected with qRT-PCR. CCK-8 was used to detect the changes in drug resistance of Hela cells. Western blotting was used to detect the expression of Sema4C in Hela cells.Results MiR-31-3p was low expressed in cervical cancer cell Hela (P < 0.05). Compared with the control group, the resistance of Hela cells transfected with miR-31-3p mimics to cisplatin was significantly reduced. MiR-31-3p can directly bind to the Sema4C gene and regulate the expression of Sema4C. Up-regulating the expression of miR-31-3p can cause the expression of Sema4C to decrease. Up-regulating the expression of Sema4C can rescue the effect of up-regulating miR-31-3p.Conclusion MiR-31-3p targets the expression of Sema4C to reduce the cisplatin resistance of cervical cancer cells.