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

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);2μmol/L、3μmol/L、4μmol/L、5μmol/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。

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.